Acta Veterinaria et Zootechnica Sinica

Research on Population Genetic Variation of Three SNPs from SLC6A14 Gene in Eleven Pig Populations

YANG Guangli;REN Jun;ZHANG Shuhong;LIU Mengzhou;ZHANG Zhiyan;HUANG Lusheng

2010, 41(1): 1-9. doi:

Abstract ( 908 )

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The aim of this research was to understand the population genetic variation and relationship, genetic differentiation between populations. In this study, 4 SNPs were identified in the porcine Solute Carrier Family 6 Member 14（SLC6A14）by directly sequencing DNA pools from 19 founder animals（2 White Duroc boars and 17 Erhualian sows）of a White Duroc × Erhualian resource population. The three mutation sites（c.1438G>A, g.7944A>T, g.21063G>T）were chosen and used for detecting the polymorphism in 11 pig breeds by PCRRFLP method. The results showed that 11 pig breeds presented varation at the g.7944A>T and g.21063G>T sites in the SLC6A14. Moreover Huai and Erhualian pigs have dominant varation at g.7944A>T site in the SLC6A14（0.01<P<0.05）, while Licha Black, Huai,Yushan Black, Hezuo Tibet, Bamei pigs also have significant varation at g.21063G>T site in the SLC6A14（P<0.01）, AT was dominant genotype. No variation was detected in the European pig populations (Landrace, Large White and Duroc). only GG genotype was presented. Whereas variation was presented at the c.1438G>A site of 8 Chinese native pig breeds. The genetic distance of 11 pig populations were calculated from 3 sites polymorphism data. The cluster tree was constructed by UPMGA method. The results showed that Chinese local pig populations had distinct genetic differentiation with exotic pig populations. It was concluded that Chinese indigenous pig breeds have higher genetic variation and diversity than the exotic pig breeds at the three SNPs, genetic differentiation existed between Chinese pig populations and exotic breeds. The majority of pig breeds were consistent with the geographical distribution and breed characteristic.

Production of Interspecies Duck/Chicken Transgenic Chimeras

LI Linfeng;PU Yabin;GONG Xuelian;BAI Chunyu;BAI Xiujuan;GUAN Weijun

2010, 41(1): 10-15. doi:

Abstract ( 858 )

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In this study, plasmid pEGFPN3 was introduced into the duck PGCs via lipofectamine in vitro. The transfected PGCs were injected into the subgerminal cavity of the recipient chicken to find a kind of method which would produce interspecies duck/chicken chimeras and contribute to transgenic study via chimeras. The Beijing duck was used as the donor and the Beijing Fatty chicken was the recipient, the duck/chicken chimeras were constructed via Windowing technique. The exogenous gene was expressed in PGCs transfected for 6 h later, and cultured for 24 h in vitro, the transfection rate reached 33.6%. 33 chicken embryos of 120 chicken eggs were obtained. Eventually 13 chimeric embryos were obtained, but there was no a surviving chicken. The exogenous gene were detected in 10.8% (13/120) of the embryos examined. Heterosexual cells were found in the gonads of 8 chimeras through PCR to amplify the Wspecific repeating sequences. The results indicated that it was possible to produce transgenic avians by constructing chimeras with the method adopted in this study. Furthermore, these results suggested that duck PGCs could migrate, colonize and proliferate in chicken gonads together with chicken′s PGCs during development.

Screening of Differentially Expressed Genes in Mammary Gland of Xinong Saanen Dairy Goat at Middle and Late Lactation Stages

WU Huijuan;LUO Jun;ZHAO Wangsheng;WANG Wei

2010, 41(1): 16-21. doi:

Abstract ( 789 )

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The objective of this study was to use suppression subtractive hybridization (SSH) to screen the differentially expressed genes in mammary gland of Xinong Saanen Dairy goats at the middle and late lactation stages, and the expression abundance of several genes were analyzed by realtime quantitative PCR (QPCR) to explore the gene expression mechanism of mammary gland in the different lactation stages of dairy goats. The subtracted cDNA libraries (ML and LM) were constructed by using mRNA of mammary gland of Xinong Saanen Dairy goat at the middle and late lactation stages as the Tester and/or Driver, clones were randomly selected and sequenced for sequence alignment, the expression abundance of several genes in mammary gland was analyzed. 30 randomly selected clones from the constructed cDNA subtraction libraries of mammary gland at middle and late lactation stages were sequenced, and the differentially expressed genes related with physiological process including cell apoptosis, antioxidation, lipid and energy metabolism etc. were obtained. Five positive clones were observed from 6 genes assayed by quantitative PCR, the expression levels of which were 1.35.5 folds higher in mammary gland of either middle or late lactation stage. The results showed that the forward and reverse subtracted cDNA libraries of mammary gland at middle and late lactation stages were constructed using SSH, the screening of 24 differentially expressed genes laid the foundation for the further studies on gene regulation in the mammary gland of dairy goats.

Study on Polymorphism of Exon9 of ABCG2 Gene in Chinese Holstein Dairy

LU Guobin;LI Hongbin;WEI Caihong;DU Lixin;LI Yurong;LU Jian;LV Yanfei

2010, 41(1): 22-26. doi:

Abstract ( 1196 )

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This study was conducted to analyze the polymorphisms of ABCG2 gene in Chinese Holstein dairy and aid to provide a theoretical foundation for further research on correlation between ABCG2 gene and milk production performance. The polymorphism of exon9 of ABCG2 gene was detected by PCRSSCP, and the SAS 9.0 was used to analyze the variance between genotypes and milk production performance. The result showed that a single nucleotide change (A/G) in exon9 resulted in the mutation from Tyrosine367 to Cysteine (Y367C) in the ABCG2 (ATP binding cassette, subfamily G, member 2) gene. There were two alleles: A and B, and their allele frequencies were 0.86 and 0.14, respectively. Three genotypes (AA, AB and BB) were found in our materials. The frequency of AA, AB and BB were 0.73, 0.26 and 0.01, respectively. The PIC (Polymorphism Information Content) was 0.22. The milk production performance, individuals with BB genotype were all highest than those with AA and AB genotypes but not significant. The relationship between Y367C and milk production performance was not obviously, but the structure of ABCG2 protein may be influenced by Y367C changed.

Screening, Identification and Tissue Expression Analysis of Septin 10 Gene Differentially Expressed in the Testis of Crossbred Bulls

LI Jiao;Chen Ruoyu;GAO Xue;LI Junya;ZHANG Lupei;ZHOU Zhengkui;CHEN Jinbao;XU Shangzhong

2010, 41(1): 27-32. doi:

Abstract ( 784 )

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This study was designed to detect differential gene expression fragments in bulls of two groups which had significant difference in scrotal circumference (SC) (P<0.01), and analyze their expression characteristics. The differential gene expression fragments were detected by differential display RTPCR, then were identified by the reverse Northern blot analysis and realtime PCR analysis, finally the expression profile of the differential gene was analyzed. One specific fragment named A71 was found. After sequencing and BLAST analysis, A71 showed high homology (99%) to bovine Septin 10 gene. The gene was expressed in various tissues, and showed significant difference between the two groups (P<0.05). The results indicated that the Septin 10 gene was differential expression gene in testicle of bulls and it might be a candidate gene in regulating reproduction traits in bulls.

Bioinformatic Analysis of TMEM66 Gene and Construction of TMEM66 Mutant Transgenic Mice

LI Xinyun;WANG Chenfang;REN Hongyan;ZHAO Shuhong;YANG Shulin;CUI Wentao;MU Yulian;LI Kui

2010, 41(1): 33-38. doi:

Abstract ( 776 )

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Transmembrane protein 66 play important roles in cell apoptosis and tumorigenesis. In order to investigate the function of TMEM66 gene, the transgenic mouse model containing the mutant TMEM66 gene was constructed in this study. The transgenic mice were generated by using microinjection method and 312 injected zygotes were implanted into 13 foster mothers. The positive transgenic mice were confirmed by PCR and Southern blotting. Also, the stability of the transgene was confirmed by passage and the way of integration of the transgene was studied using inverse PCR method. In this study, 6 positive individuals among the 55 neonatal pups were found by Southern blotting. Three positive mice were found to be able to transfer the transgene to next generation. This result indicted that the transgene was integrated into host genome stably. Inverse PCR results indicated that the transgenic DNA fragment was integrated into the host genome in a serial multicopy manner. Therefore, the transgenic mice model containing the mutant TMEM66 gene was constructed successfully. This study offered useful model for further investigation of the function of TMEM66 gene.

Effects of Dexamethasone on Absorption of Small Peptides in Small Intestine of Broilers

LI Jiaju;CAI Huiyi;LIU Guohua;CHANG Wenhuan

2010, 41(1): 39-45. doi:

Abstract ( 1117 )

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The study was conducted to investigate the effects of stress on the absorption of small peptides in the intestine of broilers. 200 21dayold male Arbor Acre broilers with similar body weight were randomly assigned into four groups and were injected subcutaneously in the abdomen with 1 mL dexamethasone (0, 0.1, 0.5 and 2.5 mg·kg1 BW) for 7 d. The body weight was measured 8 hours post the last injection on a fasting state, and the entire jejunum was collected. Histological study of jejunum was conducted, absorption of GlySar, content of plasma corticosterone, and expression of peptide transporter 1 mRNA of the broilers were analyzed. The results demonstrated that: (1)Dexamethasone reduced growth of the broilers (P<0.05)；(2) Dexamethasone inhibited absorption of GlySar by jejunum gut sacs and brush border membrane vesicles of the broilers (P<0.05);(3)Dexamethasone increased the crypt depth of the jejunum and decreased the villus height, absorption area and ratio of villus height to crypt depth (P<0.05). No significant effect on villus width was observed (P>0.05)；(4)Dexamethasone inhibited expression of peptide transporter 1 mRNA of the broilers (P<0.05). Based on this study, it could be concluded that stress inhibited the absorption of small peptides in broilers，the reasons may be related to the impairment of intestine mucosal morphology and the inhibition of expression of PepT1 mRNA.

The Effects of αKetoglutarate on Intestinal Mucosal Morphology and Function in Piglets Chronically Challenged with Lipopolysaccharide

WANG Lei;LIU Jian;HOU Yongqing;DING Binying;LIU Yulan;ZHU Huiling;LI Yongtang;WU Xin

2010, 41(1): 46-52. doi:

Abstract ( 814 )

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This study was conducted to investigate the effects of αketoglutarate (AKG) on small intestinal morphology, plasma Dxylose concentration, activities of diamine oxidase (DAO) in plasma and intestinal mucosa, and expression of mammalian target of rapamycin (mTOR) with phosphorylation (PmTOR/mTOR) in the piglets under lipopolysaccharide (LPS) challenge. Eighteen healthy crossbred (Duroc×Landrace×Yorkshire) piglets were randomly assigned into 3 treatment groups (6 replicates per group): control group, LPS group and AKG group. The control and LPS groups were fed the basal diet＋1% starch, and the AKG group was fed the basal diet＋1% AKG. On d 10, 12, 14 and 16, the piglets in the LPS and AKG groups were injected intraperitoneally with LPS at 80 μg·kg1BW, whereas piglets in the control group were injected intraperitoneally with the same dose of physiological saline. On d 16, Dxylose was orally administrated to all pigs at the dose of 0.1g·kg1BW 2 h after LPS challenge, and blood samples were collected 3 h after LPS challenge. All the piglets were sacrificed on d 17 to examine small intestinal morphology and collect intestinal mucosa for analysis. The results showed that: (1) Compared to the control group, the ratio of villus height to crypt depth in duodenum, jejunum and ileum, PmTOR/mTOR in jejunal and ileal mucosa of LPS group significantly decreased (P<0.05), while the activities of DAO in plasma significantly increased (P<0.05). (2) Compared to the LPS group, the ratio of villus height to crypt depth in the duodenum and jejunum ileum, activities of DAO in the jejunal mucosa, Dxylose content in plasma and PmTOR/mTOR in duodenal, jejunal and ileal mucosa of AKG group significantly increased (P<0.05). In conclusion, dietary supplementation with 1% AKG could improve small intestinal morphology and absorption function to some extent and alleviate the intestinal mucosal damage caused by LPS challenge in piglets, which were associated with mTOR signaling pathway.

Effects of Floor Type and Toys on Stress Response and Generic Behaviors in Growing Pigs

HU Huawei;GU Xianhong;YANG Wei

2010, 41(1): 53-59. doi:

Abstract ( 758 )

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This experiment was conducted to study the effect of floor type and toys on stress response and generic behaviors in growing pigs. Four hundreds and thirtytwo White×Landrace female growers with initial weight of (26.22±1.17) kg were randomly allotted into floor type group (no warm floor or warm floor) and toys group (no provided or provided). Every treatment had 6 replications, every replication with 18 pigs and each pen was a replication. Feeding period was 35 days. During the test period, besides collecting blood to determine the creatine kinase, blood glucose concentration, neutrophili/lymphocyte (N/L), and collecting saliva to determine the cortisol concentration, pigs′ behavior was recorded. The results showed that blood glucose level was significantly decreased in the warm floor compared with the cold floor (P=0.02). N/L ratio in the pigs provided with toys significantly decreased (P=0.04). The saliva cortisol level of warm floor group was significantly decreased in the former phase and the latter phase of the trial (P<0.01, P=0.04). Warm floor significantly decreased the frequency of comfort behavior (P=0.01), investigating behavior (P=0.02) and sidelying behavior (P<0.01). Toys significantly increased the frequency of comfort behavior (P=0.02) and disport behavior (P=0.04). In conclusion, warm floor and toys could increase the expression of generic behaviors, decrease stress and improve animal welfare.

Comparison of Immune Efficacy of Three Recombinant Fowlpox Virus Expressing HA Gene of H5 AIV

WANG Yanhong;CHEN Sujuan;LIU Wujie;QIU Xusheng;DONG Li;PENG Daxin;LIU Xiufan

2010, 41(1): 60-64. doi:

Abstract ( 764 )

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To assess the role that ORF73 or ORF214 gene of fowlpox virus(FPV) played in the regulation of immune response against FPV in commercial chickens with maternal antibodies, two recombinant fowlpox viruses (rFPV) that ORF073 or ORF214 genes inserted and coexpressed the influenza haemagglutinin (rFPVLP△73LRH5A, rFPVLP△214LRH5A) and the parental recombinant virus expressing the influenza haemagglutinin (rFPVLP12LSH5A) were constructed. These viruses were assessed for their immunological efficacy on SPF chickens and commercial chickens. The results showed that the high titer of HI antibodies was induced by three rFPV, and the protective efficacies were universal 100% in SPF chickens. However, the low HI titer was induced by rFPVLP△73LRH5A or rFPVLP△214LRH5A in commercial chickens with HI maternal antibodies up to 2.45, the protective efficacies were 16.7% and 23.3%, respectively. At the same time, rFPV with deleted ORF073 or ORF214 genes in vivo were easier to eliminate than rFPVLP12LSH5A. On the other hand, in the inoculated commercial chickens without HI maternal antibodies with rFPVLP△73LRH5A or rFPVLP△214LRH5A, the HI antibody titers increased and approached to 3.50 and 3.17, respectively after 28 days. Consequently, maternal antibodies depressed immune efficacy by rFPV with deleted ORF073 or ORF214 genes (rFPVLP△73LRH5A, rFPVLP△214LRH5A) induced.

Construction and Identification of the Recombinant Goatpox Virus with ORF8—ORF18 Deleted

ZHENG Min;LIU Qi;JIN Ningyi;SHI Kaichuang;LIANG Yuan;MO Shenglan;QU Sujie;LU Wenjun;HU Bo

2010, 41(1): 65-70. doi:

Abstract ( 750 )

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This experiment was conducted to develop a safer, high effective live goatpox virus （GTPV） attenuated vaccine and viral vector. Genome fragments of GTPV AV41 strain,including ORF7—ORF8 and ORF18—ORF19, were cloned by PCR as flanking sequences for homologous recombinant, respectively. The transfer shuttle plasmid pCFEg was constructed by inserting the expression cassettes of EGFP gene and gpt gene. The pCFEg and GTPV AV41 were cotransfected into LT cells with lipofectin, and recombinant virus was selected by fluorescence and PCR with EGFP gene primer respectively. Then, resulting recombinant virus was examined by fluorescence, PCR and TCID50 during passaging to 10th in LT cells. A recombinant GTPV with ORF8—ORF18 deleted was obtained and named as vCFEg. It was stable, and shared same multiplication ability with its parental virus (AV41) in LT cells. It suggests that genomic region of ORF8—ORF18 may be nonessential regions for replication.

Construction of Phage Display Libraries of Classic Swine Fever Virus E2 Gene and Identification of E2 Protein Epitopes

XU Hemin;WANG Qin;XU Lu;FAN Xuezheng

2010, 41(1): 71-76. doi:

Abstract ( 867 )

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The phage display peptides libraries of Classic Swine fever virus （CSFV）E2 gene were constructed to identify the potential epitopes of E2 protein by biopanning. The E2 genes of Shimen strain (SM) and hog cholera lapinized virus (HCLV) strain were cloned into T7seclect415b vector respectively. So the E2 gene phage display peptides libraries of Shimen strain and HCLV strain were constructed and named as SME2 libraries and HCLVE2 libraries. CSFV polyclonal antibody(PcAB) and seven strains of McAbs were used to screen the epitopes in SME2 libraries and HCLVE2 libraries by biopanning and phage in situ hybridization. After analyzing the sequences of selected phage recombinants, five sequences which were highly homologous with TAVSPTTLR, YYEP, TTWKEYSH, GGQ（V）VK and PDGLPHY were selected by the MAbs and CSFV PcAB. These results proved that these three sequences of TAVSPTTLR, YYEP and TTWKEYSH are dominant epitopes on E2 protein, which were reported in the previous papers. In addition, two sequences of GGQ（V）VK and PDGLPHY corresponds to the epitopes predicted by computer. So these two sequences might be the potential epitopes of E2 protein.

Development and Application of DNA Microarray for Simultaneously Detecting Actinobacillus pleuropneumoniae, Mycoplasma hyopneumoniae and Pasteurella multocida

XIAO Guosheng;;CAO Sanjie;HUANG Xiaobo;WEN Xintian

2010, 41(1): 77-85. doi:

Abstract ( 804 )

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The study aimed to establish a DNA microarray to simultaneously detecting Actinobacillus pleuropneumoniae, Mycoplasma hyopneumoniae and Pasteurella multocida. 7 different specific target DNA fragments designed and amplified from the genomes of the bacteria were used to construct the DNA microarray. The concentrations of target DNA and probes, hybridization temperature, reproducibility, specificity and sensitivity of the microarray were tested. The microarray could be used repeatedly and possessed high specificity. Microarray hybridization patterns of A. pleuropneumoniae, M. hyopneumoniae and P. multocida could be distinguished from other 9 pathogens species, such as A. lignieresii, Haemophilus parasuis, M. hyorhinitis. The sensitivity of the microarray was determined to 1050 pg of genome DNA in 50 μL labeling reaction system. 44 different serotype reference, isolates and vaccine strains of A. pleuropneumoniae, M. hyopneumoniae and P. multocida were detected with the constructed microarray. Their signal intensities and signal to noise ratio (SNRs) were greater than or equal to 1 000 and 6, respectively. Selective cultures of samples from disease pigs (n=45) and clinical healthy pigs (n=97) were detected by the microarray. Recall ratios of P. multocida, A. pleuropneumoniae, M. hyopneumoniae from disease and healthy pigs were 71.1% and 49.5%, 42.2% and 26.8%, 20% and 227%, respectively. Mixed infection rates were 42.2% and 24.7% in disease and healthy pigs respectively. The coincidence ratios between the microarray and PCR were 97.8%100%, and the ratios between the microarray and isolation method were 87.6%95.6%, in detecting clinical samples. Results indicated that the microarray have high specificity and sensitivity, and is a new tool for detection and identification of A. pleuropneumoniae, M. hyopneumoniae and P. multocida.

Effect of Dietary High Copper on Tissue Structure and BiochemicalParameters of Kidney in Chickens

CUI Wei;LI Min;PENG Xi;DENG Junliang;CUI Hengmin

2010, 41(1): 86-91. doi:

Abstract ( 860 )

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The experiment was conducted with the objective of examining the effect of dietary high copper on the renal tissue structure and biochemical parameters in chickens. 360 onedayold Avian chickens were randomly divided into six groups and feed on diets as follows: Controls (Cu 10.89 mg·kg1) and high copper groups (Cu 100 mg·kg1, high copper groupⅠ; Cu 200 mg·kg1, high copper groupⅡ; Cu 400 mg·kg1, high copper group Ⅲ; Cu 600 mg·kg1, high copper group Ⅳ; Cu 800 mg·kg1, high copper groupⅤ) for six weeks. Injuries were appeared in high copper groups Ⅲ, Ⅳ and Ⅴwhen compared with the control group, there were granular degeneration and vacuolar degeneration in the renal tubule epithelial cells. And, the renal copper contents and serum copper contents were much higher in high copper groups Ⅲ, Ⅳ and Ⅴ than that in control group(P<0.01). Also, in high copper groups Ⅲ, Ⅳ and Ⅴ, the renal malondialdehyde and hydroxy radical contents were increased (P<0.01), and the renal cuprozincsuperoxide dismutase(CuZnSOD) activities were decreased (P<0.01) compared with those of control group. The result showed that dietary copper exceed 400 mg·kg1 caused pathological injures and the impaired antioxydant function in kidney, and finally impaired renal function.

The Isolation and Identification of Neonatal Swine Intestinal Epithelial Cells

WANG Jing;ZHANG Yanming;TONG Gang;LIU Fangning;ZHOU Hongchao;HE Lei;YANG Xiaoyun;XU Yanzhao;HONG Haixia

2010, 41(1): 92-98. doi:

Abstract ( 895 )

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To provide convenience for the research of cells immortalization and viral and bacterial diseases in pig intestine, different dissociation agents and digestion protocol were used to identify the best method to isolate and culture intestinal epithelial cells. The result showed that explant culture used in this study was the best one to isolate epithelial cells from intestine of neonatal swine. The viable epithelial cells have a good capacity of proliferation. IEC attached readily to culture dishes within 22 h, proliferated significantly with 6—7 d, and reached confluence as patches with 10—12 d, lost the characteristic of epithelium after 12 generations. Morphologically features such as polygonal, oval and nonoverlapping cell was found in first 10 generations, which demonstrated the ability of the cultured cells to restore an epithelial like monolayer,and were out of shape in 11th or 12th generation. The isolated cells were identified as cells of epithelial origin by using antibodies against cytokeratine 18. Electron microscopy analysis of the ultrastructural characteristics of primary cultures of swine intestinal epithelial cells showed the presence of a wellorganized brush border and tight junction. Swine intestinal epithelial with a good capacity of proliferation and serial passage can be obtained by explant culture.

Research on the Hepatic Apoptosis and Fas mRNA Expression Induced by Manganism in Cocks

TANG Hongpeng;WANG Qiaohong;LV Zhaohui;ZHANG Liming;XU Shiwen

2010, 41(1): 99-104. doi:

Abstract ( 733 )

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The aim of this study was to investigate the effect of manganese on the hepatic apoptosis in cocks. 400 fiftyday old Hyline brown cocks were randomly divided into four groups. The 0, 600, 900 and 1 800 mg·kg1 MnCl2 were respectively added into the basic forage to establish the subchronic manganism model. After 30, 60 and 90day treatment, the livers in every group were collected to observe hepatic morphological change and detect hepatic apoptosis index and the transcription level of Fas mRNA. Results showed that among 90day treatment groups, the liver form the highest dose group took on the typical morphologic changes of apoptosis compared with the corresponding control group, and the number of the apoptotic cells gradually increased with the increase of the manganese dose. The transcription level of Fas mRNA also took on the doseeffect relationship at different treatment time.

Proteomic Analysis of Nuclei of Mammary Tissue from Healthy Cows and Clinical Mastitic Cows

WANG Jianfeng;ZHAO Xingxu;ZHANG Yong;XU Tieshan

2010, 41(1): 105-111. doi:

Abstract ( 1067 )

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The subcellular proteomics play an important role in study of minor proteins, which participating the intracellular metabolism. Subcellular proteomics can increase the minor proteins separation by twodimensional electrophoresis (2DE). A comparative analysis of proteomic profiling was conducted between nuclei of healthy mammary gland and infected clinical mastitis of dairy cow, to find more valuable biological molecular labels involved in mastitis pathogenesis. In this study, nuclei were isolated and purified by ultracentrifuge, and then twodimensional electrophoresis was applied to separate nuclear proteins. Imagine analysis for selection of differentiallyexpressed protein spots was conducted by PDQUest 7.4 software. By ion trap mass spectrometry equipped with surveyor HPLC system proteins were identified. 22 spots from electrophoresis gels were selected as differentiallyexpressed proteins for MS analysis and 17 proteins were identified. Seven of differentiallyexpressed proteins showed upregulated expression and 2 proteins showed downregulated expression during mastitis. 5 proteins only expressed in healthy mammary gland and 3 proteins only expressed in mammary gland which infected with clinical mastitis. The differentiallyexpressed proteins separated in this study had a wide range of functions which are related to the formation of cellular skeleton, metabolic regulation and apoptosis regulation. These suggested that nuclei of mammary gland had experienced fundamental changes in structure and metabolism when contracted clinical mastitis.

Immunization of Mother Mice with Recombinant Myostatin Cdomain Increasing the Body Weight of Neonatal Mice

MA Yi;LI Qing;CHEN Xiaoqiang;WANG Wenjie;GUAN Hong;AN Xiaorong;CHEN Yongfu

2010, 41(1): 112-116. doi:

Abstract ( 1111 )

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The potential of using recombinant MSTN as vaccine to improve muscle mass was explored in this study. Firstly, the recombinant myostatin Cdomain was obtained through prokaryotic expression and purification. Then female Kunming White mice after immunization with recombinant Cdomain were mated with male mice. The indirect ELISA assay was used to detect the titer of antibody against MSTN in mother mice. The body weights of mice were evaluated each week. The results showed: (1) The body weight of mother mice in experiment group didn′t change significantly compared with that in control group (P>0.05). But during the late pregnancy (the 11th week after immunization), the increase of body weight of experiment group exceeded significantly that in control group (P<0.05). The body weight was (56.23±3.37)g in experiment group，and (54.11±3.18)g in control group. In the 12th week after immunization, the body weight of experiment group was (79.16±3.72)g, and that of control group was (7523±344)g. The former was significantly higher than the latter (P<0.01). After the delivery, the difference of body weight in two groups disappeared again (P>0.05); (2) The body weight of neonatal mice in experiment group was 25.8% higher than that in control group (P<0.01), but this difference decreased with the development of neonatal mice. These results proved that immunization of mother mice with recombinant MSTN Cdomain protein could significantly increase the body weight of neonatal mice.

The Antioxidation Protective Effects of Vitamin E on Frozenthawed Semen in Qinchuan Cattle

ZHAO Xianlin;ZAN Linsen;HU Jianhong;GUI Linsheng;HAO Ruijie

2010, 41(1): 117-122. doi:

Abstract ( 1146 )

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The aims of this study were to determine the antioxidation protective effects of vitamin E on the quality of straw frozen semen and antioxidant enzyme activity in seminal plasma of Qinchuan cattle. In this experiment, the vitamin E with different concentrations (0.00, 002, 004, 0.06, 0.08, 0.10 mg·mL1) were added in the ordinary dilution. After thawing the straw frozen semen(37.5 ℃,30 s), the motility of sperm, the integrity rate of sperm acrosome and other parameters were analyzed by WEILI sperm analysalor. Then the activity of SOD, GSH, GR and CAT were detected by the related kit. The results showed that when the addition of vitamin E was 0.04 mg·mL1, there were significant differences between experiment and control groups in motility of sperm (P<0.05) and also had significant differences in enzyme activity of SOD, GSH and CAT (P<0.05). When the addition of vitamin E was 0.06 mg·mL1, the motility of sperm were significant higher than that of control group, the integrity rate of sperm acrosome was significant higher than that of control group (P<0.01); there were significant differences between experiment and control groups in the activity of GR and CAT. When the vitamin E concentration was 0.08 mg·mL1, the sperm motility, the motility and the integrity rate of acrosome had very significant differences (P<0.01) between experiment and control groups, there were significant differences in GR and CAT activity between experiment and control groups. With the addition of vitamin E in the dilution of frozen semen, the quality of frozenthawed semen and enzyme activity of SOD, CAT, GSH and GR in seminal plasma of Qinchuan cattle can be effectively improved, and the appropriate additions of vitamin E in ordinary dilution were 0.040.08 mg·mL1.

The Expression of Bovine βDefensin 5 Gene in the Pichia pastoris

LI Hongbin;LONG Jing;DU Lixin

2010, 41(1): 123-126. doi:

Abstract ( 773 )

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Based on the peripheral blood leukocyte cDNA library constructed by suppression subtractive hybridization (SSH) in our lab, bovine βdefensin 5 gene (BNBD5) was chosed as one of the candidate genes. The aim of the current research was to look for molecular events and establish fundamental materials and techniques for further study. Finally, the coding sequence (CDS) of BNBD5 gene was successfully amplified. The Pichia pastoris expression vector was constructed and the BNBD5 protein was expressed. This research lays the foundation for preparation of antibodies of BNBD5.

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