Acta Veterinaria et Zootechnica Sinica

Polymorphism Analysis on Exon 14 of Mx1 Gene in Wild Boar and 16 Native and Exotic Pig Breeds

WU Sheng-long;BAO Wen-bin;JU Hui-ping;DING Hao;LI Bi-chun;ZHU Guo-qiang;CHEN Guo-hong

2008, 39(3): 257-261. doi:

Abstract ( 1307 )

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By the PCR-RFLP method, the polymorphisms of exon 14 of Mx1 gene were detected in wild boar and 16 native and exotic pig breeds, with the analysis of restrictive enzyme Hin6Ⅰdigestion, there were 6 genotypes and 3 alleles. The exotic pig breeds and Songliao Black pigs appeared AA, AC and CC genotypes, all other Chinese native pig breeds appeared AA, BB and AB genotypes, the Sutai pigs appeared all six kinds of genotypes. Among all pig breeds in this study, the allele B only appeared in Chinese native pig breeds, while the allele C only appeared in exotic pig breeds and developed pig breeds which originated from exotic pig breeds. Dendrogram of phylogenetic relationships among these pig breeds based on polymorphism of Mx1 gene also showed that genotype distribution in Mx1 exon 14 was obviously different between Chinese native pig breeds and exotic pig breeds, all exotic pig breeds and two developed pig breeds formed one cluster, wild boar and all Chinese native pig breeds were clustered in the other large population, among this population, Fengjing pig, Erhualian pig and Meishan pig formed one branch firstly, and it had the nearest genetic relationship with wild boar. The frequency of AB genotype was relative higher in the individuals of wild boar and Tibetan pig, which were 0.515 and 0.302, respectively. The frequency of AB genotype in the populations of Fengjing pig, Erhualian pig and Meishan pig ranged from 0.348 to 0.591. All these results indicated that the individuals with AB genotype maybe have stronger ability of virus-resistance, and the Taihu pigs (Fengjing pig, Erhualian pig and Meishan pig) were perhaps the good potential materials for virus-resistant breeding in the future.

Genetic Co-adaptability of Structural Genes in Wild Boars Populations

LI Xiao-lin;PAN Qing-jie;MIN Ling-jiang;SHEN Wei;LIU Di

2008, 39(3): 262-267. doi:

Abstract ( 1499 )

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Total 42 Northeast wild boars derived from Jilin、Heilongjiang and Liaoning provinces were investigated, and the genetic polymorphism of 7 structural sites in wild boars populations were detected using the techniques of PCR-SSCP and PCR-RFLP. Subsequently, the genetic co-adaptability of these genes was analyzed and the results showed that 5 of 7 sites (71.43%) are genetic disequilibrium, 6 of 18 sites combinations in different chromosomes are genetic disequilibrium for genetic coadaptability. All of 3 sites combinations in the same chromosome are genetic disequilibrium. Leptin-ECF18R, however, is in linkage disequilibrium. The cause of the genetic equilibrium in the sites combination of Leptin-ECF18R maybe the opposite effect between genetic coadaptability and linkage disequilibrium.

Association of the Polymorphism in Porcine Aquaporin 7 Gene with Fat Deposition

WANG Li-xia;XIONG Yuan-zhu;ZUO Bo

2008, 39(3): 268-272. doi:

Abstract ( 926 )

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Aquaporin 7 (AQP7) is an newly discovered aquaglyceroprotein and responsible for the movement of glycerol In this study, we firstly obtained the sequence of intron 4 of the porcine AQP7 gene from Large White, Landrace and Meishan pig Sequence comparison revealed a G/A mutation which was detected by PCRPstIRFLP The G allele was predominant in the Large White and Landrace pigs, while there was higher A allele frequency in Meishan pigs Association analysis between the polymorphism and fat deposition traits was performed in 234 Large white×Meishan F2 pigs Significant associations of the genotype with fat deposition traits such as leaf fat weight (LFW), internal fat rate (IFR), backfat thickness (BT), fat percentage (FP) and ratio of lean meat versus fat meat (RLF) were found The individuals with GG genotype have lower fat deposition value than those with AA genotype and allele G was significantly associated with the decreasing of fat deposition

Tissue Differential Expression of PPAR-γ in Bamei Pig

WANG Bo;WU Jiang-wei;YANG Gong-she

2008, 39(3): 273-277. doi:

Abstract ( 878 )

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Total RNA and protein were extracted from visceral adipose tissue, subcutaneous adipose tissue, skeletal muscle, kidney, liver, heart, lung and spleen of Bamei pig. The primers of PPAR-γ1 and PPAR-γ2 were designed and synthesized. The specific expression of PPAR-γ1 and PPAR-γ2 in different tissues were investigated by Semi-Quantitative RT-PCR and Western Blot. The results showed that the mRNA and protein of PPAR-γ1 were expressed in all abovementioned tissues, and highly expressed in visceral adipose tissue，subcutaneous adipose tissue and lower expressed in lung; PPAR-γ2 mRNA and protein were highly expressed in visceral adipose tissue, subcutaneous adipose tissue, spleen and heart, while remarkable lower in lung, kidney, liver and muscle. In conclusion, the results suggested that PPAR-γ might have various effects on different tissues of pig.

Identification of Single Nucleotide Polymorphisms of SCD and Sirt1 and Evaluation of Their Genetic Diversities in Chickens

LIU San-feng;REN Jun;GUO Yuan-mei;YANG Yan;ZHANG Zhi-yan;HUANG Lu-sheng

2008, 39(3): 278-285. doi:

Abstract ( 837 )

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StearoylCoA desaturase (SCD) and Sirtuin 1 (Sirt1) on the chicken chromosome 6 are two functional genes that are associated with fat deposition In this study，single nucleotide polymorphisms (SNPs) were screened by comparative sequence analysis using DNA pool of three divergent chicken breeds including Taihe Silk, White Recessive Rock and Xinghua A total of 13 SNPs were identified in the SCD gene, of which 5 SNPs were in the coding region (2 missense mutations) and 10 SNPs were novel SNPs And 35 SNPs were identified in the Sirt1 gene including 10 SNPs in the coding region (2 missense mutations), 3 InDels and 18 novel SNPs The SCD c492 G>A and the Sirt1 c1753 G>A polymorphism were genotyped in six chicken breeds (White Recessive Rock, Xinghua, Taihe Silk, Baier Huang, Kangle, Nancheng Black) using single strandedconformation polymorphism (SSCP) assay The results showed that the genetic polymorphisms were abundant, providing new markers for further association analysis in this genomic region

Study on Genetic Diversity and Genetic Differentiation of Indigenous Geese Breeds in Eastern China

CHEN Kuan-wei;SONG Wei-tao;XU Wen-juan;LI Hui-fang;TANG Qing-ping

2008, 39(3): 286-290. doi:

Abstract ( 942 )

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The genetic diversity and genetic differentiation of 14 indigenous geese breeds in Eastern China were surveyed using 25 microsatellite markers. Using polymorphism information content (PIC), mean heterozygosity (H), number of effective alleles, DA genetic distances, gene flow (Nm) and Fstatistics, we evaluated the genetic diversity and genetic differentiation. The results showed that the number of alleles per locus ranged from 4 to 16. All populations showed high levels of heterozygosity, the lowest was Yane geese (0.566 2) and the highest was Baizi geese (0.774 8). Considerable genetic differentiation was observed and 27.5% of the total genetic variation was from the breeds differences. Neighbour-joining dendrogrm was constructed based on the DA genetic distance, and four main clusters were found. Analyzing the correlation between genetic differentiations and geographical distances, we presumed that genetic differentiations among the 14 indigenous geese breeds may not be related to geographical distances.

Study on Genetic Structure of Domestic Duck Populations in Middle China

LI Hui-fang;LI Bi-chun;YANG Ning;CHEN Kuan-wei;TANG Qing-ping;SONG Wei-tao

2008, 39(3): 291-295. doi:

Abstract ( 1321 )

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By using microsatellite markers, the genetic structure of six domestic duck resources distributed in Middle China, including Huainan Sheldrake, Youxian Sheldrake, Linwu, Jingjiang Sheldrake, Mianyang Sheldrake and Enshi Sheldrake, was analyzed in this experiment. The results showed that the mean heterozygosity of six duck populations was 0.580 1, the highest of which was Jingjiang Sheldrake (0.551 2), and the lowest was Youxian Sheldrake (0.603 5). The value of heterozygosity reflected the rich diversity of those populations. And considerable population differentiation was observed by F-statistics analysis and 22.5% of the total genetic variation came from population differences, this result firmly affirmed each breed was of its own genetic diversity. The DA genetic distances suggested the longer differentiation existed among those breeds. Based on the distances among those populations, the results of NJ clustering were formed, the first group was Jingjiang Sheldrake, Mianyang Sheldrake and Enshi Sheldrake; the second group was Youxian Sheldrake and Linwu; the third group was Huainan Sheldrake. In terms of the results, those could provide not only basic molecular data for the research on the characteristics of local breeds in Middle China but also scientific basis for the conservation and utilization of those breeds.

Effects of Bcl-2 Gene Interference on the Apoptosis, Proliferation and Progesterone Secretion of Goose Follicular Granulosa Cells

CHEN Xiu-ping;CHEN Feng-jian;JIANG Xun-ping;LI Zhao-guo;DING Jia-tong

2008, 39(3): 296-301. doi:

Abstract ( 1320 )

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According to the published sequences, three siRNAs (Small interference RNA) were designed, and expression vectors were constructed and transfected into goose granulosa cells cultured in vitro. The expression level of Bcl-2 protein, apoptosis and proliferation of granulosa cells were analyzed by flow cytometry 48 h after the transfection, and the progesterone (P) in the medium was assayed using RIA. Meanwhile, apoptosis indexes and Bcl-2 protein level of follicular granulosa cells with different rank were also analyzed. Our results indicated that: (1) Levels of Bcl-2 protein in follicular granulosa cells with different rank were different, the highest expression level was observed in the smallest preovulatory follicle (SPY), and levels of Bcl-2 protein in the healthy follicles were significant higher than that in atresic follicles (P < 0.05); (2) Bcl-2 protein levels of test groups were significant lower than that of control (P < 0.05), while apoptosis indexes (AI), proliferation indexes (PI) and P secretion of test groups were higher than that of control (P < 0.05).

Determination of Plasma Melatonin of Female Yaks with Different Reproductive Rhythms by Reversed-phase High-performance Liquid Chromatography in Warm and Cold Seasons

WEN Yong-li;WANG Yong;QI Sharina;WANG Yan-hong;MA Li

2008, 39(3): 302-308. doi:

Abstract ( 1526 )

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The differences were studied among seasons, reproductive rhythms, breeding active and resting terms. The plasma concentrations of melatonin of adult female yaks were determined by reversed-phase high-performance liquid chromatography. A total of 63 adult female yaks were selected as experimental animals and divided into three groups: A with calving every year and breeding active term, B with calving per two years and full milking and breeding resting term, C with calving per two years and semimilking and breeding active term. Each group had 21 yaks. In the sampling period of July 21-23 in warm season and November 21-23 in cold season, the yaks were exposed to 12.00 L∶12.00 D and 14.65 L∶9.35 D, respectively. Blood samples were collected for 50 minutes starting at 2 h before sunset. A quantity of 20 mL jugular vein blood was drawn from every animal and anticoagulated with heparin sodium. Plasma obtained after centrifugation was estimated for melatonin. The content of plasma melatonin of group A ((68.90±5.82) pg/mL) was higher than that of group B ((44.38±2.61) pg/mL) (P<0.01) and group C ((57.51±3.82) pg/mL) (P<0.05). The content of plasma melatonin in cold season was lower than that in warm season (P<0.05). The level of plasma melatonin of yak cows in reproductive active term was higher than that of it in fallow term (P<0.05). There was a seasonal rhythm in the secretion of plasma melatonin of yak cows. The concentration of plasma melatonin had an effect on the seasonal breeding and reproductive rhythms of female yaks.

Recombinase RecA-mediated Mouse Transgenesis by Microinjection

ZHAO Yong-zhen;XIAO Bang;ZHANG Xing-ju;TANG Zhong-lin;CUI Wen-tao;YANG Shu-lin;LI Kui

2008, 39(3): 309-313. doi:

Abstract ( 832 )

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One main reason of low efficiency producing transgenic animal by microinjection is digestion of foreign DNA by nucleases in nuclear. Recombinase RecA can protect DNA from digestion by phosphodiesterases or nucleases. A recombinase RecA-mediated mouse transgenesis method was developed. Further, we discussed the effect of recombinase RecA on production efficiency of transgenic mouse by microinjection. 41 F0 mice were obtained by microinjecting complexes of single strand DNA and RecA protein. Using single strand DNA and double strand DNA for microinjection, 28 and 32 F0 mice were obtained, respectively. All F0 mice were determined to be transgenic by PCR and Southern blot. The result showed that 14.6%（6/41） of F0 mice produced by pronuclear microinjection of RecA coated ssDNA was transgenic. The production efficiency of transgenic mouse was 6.2% when dsDNA was used. There was no transgenic when ssDNA was used.

Effect of Xylanase on Intestinal Amino Acids Transporters mRNA Expression of Yellow-feathered Chicken Fed Wheat-based Diet

FENG Ding-yuan;TAN Hui-ze;WANG Xiu-qi;ZOU Shi-geng;ZUO Jian-jun;HUANG Yan-hua;DONG Ze-min;ZHANG Chang-ming;YE Hui

2008, 39(3): 314-319. doi:

Abstract ( 1530 )

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480 healthy 1-day-old male parent yellow-feathered chicken were assigned randomly into A and B groups, and each group had 6 pens (40 birds/pen). Group A were fed with wheatbased diet and Group B were fed with wheat based-diet with xylanase( 1.2×10⁴ U/kg diet). On day 16, two birds (closest to the mean weight) were selected from each replication. Sample of jejunum and ileum was collected for the determination of mRNA expression of cationic amino acid transporters. Xylanase significantly increased the expression abundance of rBAT and CAT4 mRNA in the jejunum of yellow-feathered chicken fed the wheat-based diet (P<0.05) and had the trend to increase the expression abundance of y+LAT2 and CAT1 mRNA in jejunum (P>0.05). It had no effect on the expression of rBAT mRNA in ileum but had the trend to increase the expression abundance of y+LAT2, CAT1 and CAT4 mRNA in ileum (P>0.05).

The Effect of Parity on Conjugated Linoleic Acid (CLA) Content in Milk Fat and CLA-Desaturase Index from Holstein Dairy Cows

HOU Jun-cai;LIU Yan-ping;HUO Gui-cheng;REN Da-xi;TI Wei-gang

2008, 39(3): 320-326. doi:

Abstract ( 1112 )

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The milk samples of two hundred and thirty-five lactating Holstein dairy cows in north China were selected. In order to avoid confounding effects of diet and season, Holstein cows were fed a single diet and milk sampled on the same day. The performance of lactating dairy cows and composition of the fatty acids in milk fat were measured to investigate the effect of parity on conjugated linoleic acid (CLA) in milk fat and △⁹-desaturase indexes from Holstein dairy cows.The results showed that the average content of cis-9, trans-11 CLA was 5.14 mg/g fatty acids in milk fat, and the variation range was 1.5-10.5 mg/g fatty acids. The average content of cis-9, trans-11 CLA of primiparous was 5.19 mg/g fatty acids in milk fat, while it was 5.11 mg/g fatty acids of multiparous, and there's no significant difference between them（P>0.05）. There was significant difference between the △⁹-desaturase index of cis-9 C14∶1 and cis-9 C16∶1 of primiparous and multiparous（P<0.05）, and there was no significant difference between the △⁹-desaturase index of cis-9 C18∶1 and cis-9, trans-11 CLA of primiparous and multiparous（P>0.05）.

Construction and Protective Efficacies of Two Recombinant Fowlpox Viruses Expressing the HA Gene of H9 Subtype AIV

LIU Wu-jie;SUN Lei;CHEN Su-juan;XU Zhong-lin;LIU Jin-biao;LIU Xiu-fan

2008, 39(3): 327-332. doi:

Abstract ( 1502 )

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The HA gene from subtype H9N2 AIV was directionally inserted into the vector pP12LS, resulting in transfer vector pP12LSH9A. Then the pP12LSH9A was used to transfect the chicken embryo fibroblast cells, which were preinfected with strain 282E4 or LP of FPV, respectively. After serial blue plaque screening, we obtained two purified rFPVs: rFPV₂₈₂-12LSH9A and rFPV_LP-12LSH9A. SPF and antibody positive commercial chickens were immunized with the two rFPVs to investigate their protective efficacies. Chickens immunized with homologous AIV inactivated vaccine or widetype FPV served as controls.The protective efficacies of rFPV₂₈₂-12LSH9A and rFPV_LP-12LSH9A in SPF chickens were both 100%，those ones in commercial chickens were 88% and 92%, respectively. The results revealed that the rFPV₂₈₂-12LSH9A and rFPV_LP-12LSH9A are two effective candidate vaccines against H9 subtype AIV even in the presence of maternal antibodies against H9 subtype AIV.

Rescuing of Foot-and-mouth Disease Virus O/LZ Strain

MA Ming-xiao;JIN Ning-yi;SHEN Guo-shun;LIU Hui-juan;HUO Xiao-wei;JIN Ming-lan;ZHENG Min;LU Hui-jun;JIN Kuo-shi

2008, 39(3): 333-336. doi:

Abstract ( 1517 )

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The cloned cDNA derived from the genome of FMDV O/LZ strain was used as template and was transcripted to produce RNA with T7 RiboMAXTM Express Large Scale RNA Production System. The RNA was transfected into BHK cells. The typical cytopathic effect which were caused by rescusing FMDV was observed. Furthermore，the rescusing FMDV were verified by RT-PCR，IFA and observation of immunoelectron microscope. The results suggested that the FMDV were successfully rescued. This study have paved the way for exploring vaccines against FMDV，pathogenic mechanism and so on.

The Establishment and Application of Inhibited Expression Vector of Prion Protein by RNA Interference

LI Yu-rong;ZHOU Xiang-mei;YIN Xiao-min;YANG Jian-min;QIAO Jun-wen;ZHAO De-ming

2008, 39(3): 337-342. doi:

Abstract ( 1287 )

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In present research,we intend to construct pG-super-siPrP expression plasmid and explore its function in C6 cells．shPrP was subcloned into pG-super. Recombinant plasmid pG-super-shPrP was transformed into Top10 E.coli, and the ampicillin resistant clones were identified by PCR and DNA sequencing．C6 cells were transfected with the identified pG-super-shPrP and pG-super (the control group) by LipofectamineTM2000．PrP mRNA was quantified by realtime RT-PCR.The results showed that the expression of PrP mRNA in pG-super-shPrP group decreased by 34.2% compared with the control group (P<0.05)．The detection results of the total activity of superoxide dismutase indicated that PrP promoted the SOD activity.In conclusion, pG-super-shPrP expression plasmid was constructed effectively and took effect in C6 cells，which helped to further study about PrP biochemical and physiological role in animal and offer a attractive therapeutic approach to fight against prion disease.

Isolation of Actinobacillus pleuropneumoniae by Immunomagnetic Beads Technique

. The National Key Laboratory of Agricultural Microbiology;Huazhong Agricultural University;Wuhan 000;China;. Hubei Key Laboratory of Animal Embryo and Molecular Breeding;Wuhan 00;China

2008, 39(3): 343-348. doi:

Abstract ( 854 )

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The conservative outer membrane lipoprotein of Actinobacillus pleuropneumoniae(APP) was expressed, purified and used to immunize the rabbit for production of antiserum, from which IgG was purified and used to sensitize the immunomagnetic beads. The optimal conditions for coating immunomagnetic beads with IgG, including IgG concentration and time, were determined. The protocol for detection of App by coated immunomagnetic bead was developed. All of the 14 APP reference strains could be detected by the new method. The analytical sensitivity of the assay was higher than that of the traditional procedure. Ten strains of twenty suspected strains were identified as APP,and consistently confirmed by ApxⅣ-based PCR. The coincidence rate between ApxⅣ-based PCR and developed immunomagnetic beads separation was 100%. The challenged APP serotype 1 and serotype 7 could be recovered by the novel method. Finally, this method was clinically applied, and five APP strains were isolated from the samples that collected from Henan, Hunan and Hubei province. These isolates were confirmed by ApxⅣ-based PCR.The results indicated that the developed immunomagnetic beadbased method can be used to separate APP.

Distribution of Orexin B in the Central Nervous System of Pig

SU Juan;LEI Zhi-hai;NING Hong-mei;SONG Jie;JIA Cui-ping

2008, 39(3): 349-354. doi:

Abstract ( 848 )

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Distribution of orexin B immunoreactive neurons and fibers in the central nervous system of pigs was studied by immunohistochemistry. The results showed that orexin B positive neurons and fibers were observed in the hypothalamus, brain stem and spinal cord. In the hypothalamus, orexin B immunoreactive neurons and fibers were found in the the preoptic area, suprachiasmatic nucleus, paraventricular nucleus, periventricular nucleus, dorsomedial hypothalamic nucleus, perifornical nucleus, mammillary nucleus, lateral and posterior hypothalamic areas, and with dense distribution in the perfornical nucleus, zona incerta, dorsomedial nucleus and posterior hypothalamic area and sparse distribution in the preoptic area, lateral hypothalamic area and mammillary nucleus. In the brain stem, orexin B immunoreactive neurons were observed in the locus ceruleus, vestibular nucleus, inferior olivary nucleus, nucleus of solitary tract and nucleus ambiguus. In the spinal cord, orexin B immunoreactive neurons were sparse and distributed in the lamina Ⅳof the dorsal horn of cervical segment of the spinal cord. Furthermore, dense orexin B immunoreactive fibers were found in the diagonal band, septal area and bed nucleus of the stria terminalis. Our results provided the bases for the studies of the physiological functions of orexin B in pigs.

Observation on the Pulmonary Capillary in One-day Old Yak (Bos grunniens) under the Scanning Electron Microscope

DU Xiao-xia;YU Hong-xian;LIU Ying;DU Xiao-hua;GUO Jian-qiang;ZHAO Jian-hong;WANG Yu;LIU Dong-yang;LI Ren-ge;ZHAO Hai-tao;LIU Bin

2008, 39(3): 355-359. doi:

Abstract ( 942 )

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The present study characterized the pulmonary microvascular architecture in the parenchyma of newborn yak by vascular corrosion cast and scanning electron microscopy in order to explore the distributing patterns of pulmonary microvessels in newborn yak, the mechanisms of the microvascular growth and development during the postnatal period. We found irregular, honeycomb like alveolar capillary network on the cut surface at a low magnification. In some fields of view, the alveolar capillary networks extended incompletely, and the alveolar cavities varied in shape and size. We also found completely expanded alveoli, half expanded alveoli, slightly expanded alveoli and unexpanded alveoli. There were three different forms of alveolar capillary network, including sheetlike, sievelike and weblike network. Also the capillary network in the alveolar septa could be classed into single-layered, double-layered and adjacent capillary networks. In addition, the oriented imprints of the endothelial cell nuclei were found on the surface of the well-expanded capillaries.

Effect of Chinese Medicine Effective Component Prescription on in vitro Proliferation of Mouse Intestinal Lymphocyte after Stimulation with High Temperature

WANG Zi-li;DONG Hong;LIU Feng-hua;MAO Shuai;LIU Ren-zhi;XU Jian-qin

2008, 39(3): 360-364. doi:

Abstract ( 1438 )

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The effective components of herba agastachis, rhizoma atractylodis, gypsum fibrosum and cortex phellodendri were composed in different combination according to the principal of traditional Chinese composing prescription and orthogonal design to form 8 prescriptions. The effects of different prescriptions on in vitro proliferation of mouse intestinal lymphocyte after stimulation with high temperature were studied. The results showed that herba agastachis aetherolea and cortex phellodendri alkaloid were the major drugs in the prescription, rhizoma atractylodis aetherolea and gypsum fibrosum water extract were adjuvant drug and messenger drug. The prescription composed of 200 μg/mL herba agastachis aetherolea, cortex phellodendri alkaloid and rhizoma atractylodis aetherolea, 100 μg/mL gypsum fibrosum water extract, has the best effect on the proliferation of intestinal lymphocyte, and the interaction of herba agastachis and rhizoma atractylodis aetherolea was better.

Cloning of Small-tailed Han Sheep YB-1 cDNA and Expression in E. coli

SONG Xue-mei;LI Hong-bin;DU Li-xin

2008, 39(3): 365-367. doi:

Abstract ( 733 )

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Full-length cDNA of YB-1 gene was cloned from testicle in Small-tailed Han sheep by RT-PCR. pET32a expression system was used to express open reading frame (ORF) of YB-1 in E.coli with induction of IPTG at different concentrations. The low expression level of fusion protein revealed that IPTG concentration did not affect on its expression level. To improve the expression level of fusion protein, ampicillin was replaced by carbenicillin in cultural medium, which increased expression efficiency.

Effect of Different Photoperiods on Blood Hormone of Laying Hens

HUANG Ren-lu;CHEN Hui;DI Ke-qian;ZHANG Jing-qian;LI Jun-qiao;ZHANG Zhen-hong;PAN Dong

2008, 39(3): 368-371. doi:

Abstract ( 842 )

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At the age of 20 week, 320 Hyline (Brown) commercial hens were randomly assigned into 8 groups, with 4 replications per group, 10 hens in each replication. According to abruptly and slowly increscent methods, the experiment was designed to 11L∶13D, 13L∶11D, 15L∶9D, 17L∶7D four photoperiods. At the end of 29 week, 2 hens were randomly selected from each replication, in order to detect the level of FSH, LH, P4 and E2.The results showed that FSH cooperate with LH, which could accelerate ovum to maturate and arose ovary secreting E2 and P4, then ovulating. 11L∶13D photoperiod group by slowly increscent method(GroupⅡ) had the highest surge level of FSH and LH. The higher hormone secretion was not related to the longer photoperiod.

Detection of Distribution of Porcine Circovirus Type 2 in Experimentally InfectedPiglets with Immunohistochemical Method

TANG Ning;LV Yan-li;GUO Xin;YANG Han-chun;LAI Ting;FENG Hua-bing

2008, 39(3): 372-375. doi:

Abstract ( 881 )

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28-day-old healthy conventional piglets were inoculated nasally with porcine circovirus type 2 (PCV2) BF strain, the piglets were necropsy on different day post inoculation. Tissues were collected for preparing tissue sections and immunohistochemistry protocol were developed to detect distribution of PCV2 antigens in tissue samples. Results showed that in lymphoid tissues such as lymph node, spleen and tonsil were positive for PCV2 antigen. PCV2 positive signals also presented in heart, liver, lung, kidney, stomach, duodenum, cerebrum and cerebellum. Other tissues were negative for PCV2 antigen. Viral antigens were localized predominately in the cytoplasm of the infected cells and rarely observed in the nucleus of the cells. Results indicated that infection indeed presented in the tissues of the experimental piglets.

An Unknown Taenia Solium’ Gene of SLC10 Expressed in Pichia pastoris

WANG Ying;LUO Xue-nong;ZHENG Ya-dong;MA Quan-ying;DING Jun-tao;JING Zhi-zhong;CAI Xue-peng

2008, 39(3): 376-379. doi:

Abstract ( 819 )

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An unknown gene, SLC10, was cloned by RT-PCR from Cysticercus cellulosae on the base of transspliced leader sequence. The SLC10 gene was cloned from vector of pGEM-SLC10 and inserted into pPIC9k to construct a recombinant secretory vector named pPIC9K-SLC10. Then the pPIC9k-SLC10 was transformed into E.coli JM109.The recombinant vector was identified by PCR, enzymatic analysis and sequencing. The positive plasmid was linearized with Sac I and transformed into GS115 by electroporation and the insert was integrated into genomic DNA of Pichia pastoris by means of homologous recombination. The recombinants with multiple integrated copies of SLC10 were screened using gradient concentration G418 and induced with methanol. The results of SDS-PAGE and Western-blot showed that the expressed protein was not of immunoreaction.

Development and Application of Competitive ELISA Kit for Rapid Detection of Streptomycin Residues

FAN Guo-ying;WANG Shun-gang;WANG Zi-liang;ZHANG Gai-ping;DENG Run-guang;YANG Yan-yan

2008, 39(3): 380-384. doi:

Abstract ( 868 )

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BALB/c mice were immunized with streptomycin-bovine serum albumin and hybridoma lines that secrete monoclonal antibody against streptomycin(SM mAb) were filtered with cell fusion. A competitive ELISA test kit for detection of streptomycin（SM-Kit）was developed with anti-SM monoclonal antibody（SM mAb） and its sensitivity，veracity，specificity and the effect of matrix were tested, respectively. At the same time, the urine of the swine was detected with this ELISA-Kit. The results indicated that the SM-Kit has little or no crossreactivity with other antibiotics besides of dihydrostreptomycin；the linear detection was 1.0 to 128.0 μg/L（R2＝0.925 1），the sensitivity was 0.45 μg/L，the IC50 was 7.76 μg/L and the detection limit was 1 μg/L. The recoveries of SM spiked in pig urine was 84.1%. The matrix has no effect on the results of the SM-Kit. The SM-Kit possessed rapidity, sensitivity, specificity and briefness and was proved useful in the rapid test of SM residues in animal.

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