单核细胞增生李斯特菌非编码RNA rli87基因缺失株的构建、鉴定及其生长特性初步研究

doi:10.11843/j.issn.0366-6964.2015.06.016

Abstract

Abstract:

In this study，non-coding RNA rli87 gene deletion strain of LM was constructed and identified，and growth characteristics of deletion strain was studied.The rli87 gene deletion mutant was constructed by fusion PCR method，and then pKSV7-Δrli87 shuttle vector was constructed，which was transformed into competent cells LM-SB5.Homologous recombination was conducted using temperature (42 ℃) and chloramphenicol (10 μg•mL^-1) resistance to achieve the rli87 gene deletion strain，and growth curves of wild strain and deletion strain were assayed at 37 ℃.The PCR and sequencing results confirmed that LM-Δrli87 was successfully obtained.PCR identification results showed that the deletion strain has good genetic stability through 25 generations of continuous passage.Comparison with the wild strain，there was no significant difference (P>0.05) in growth between deletion mutant and wild strain at 37 ℃.The results suggest that rli87 gene did not play significant role in regulatory growth under the conditions of 37 ℃.

CLC Number:

S852.61

XIE Kun，QIAO Jun，MENG Qing-ling，PENG Ye-long，ZHAO Hai-long，MA Yu，CHEN Cheng，CAI Xue-peng，CHEN Chuang-fu. Construction，Identification of ncRNA rli87 Gene Deletion Strains Listeria monocytogenes and Its Growth Characteristics[J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2015, 46(6): 998-1003.

0
/ / Recommend

Add to citation manager EndNote|Reference Manager|ProCite|BibTeX|RefWorks

URL: https://www.xmsyxb.com/EN/10.11843/j.issn.0366-6964.2015.06.016

https://www.xmsyxb.com/EN/Y2015/V46/I6/998

References

［1］陆承平.兽医微生物学［M］.3版.北京：中国农业出版社，2001.
LU C P.Veterinary Microbiology ［M］.3rd.Beijing ：Chinese Agricultural Press，2001.(in Chinese)
［2］CHRISTIANSEN J K，NIELSEN J S，EBERSBACH T，et al.Identification of small Hfq-binding RNAs in Listeria monocytogenes［J］. RNA，2006，12(7)：1383-1396.
［3］IZAR B，MRAHEIL M A，HAIN T.Identification and role of regulatory non-coding RNAs in Listeria monocytogenes［J］.Int J Mol Sci，2011，12(8)：5070-5079.
［4］WANG K C，CHANG H Y.Molecular mechanisms of long noncoding RNAs ［J］.Mol Cell，2011，43(6)：904-914.
［5］高原，惠宁，刘善荣.长链非编码RNA的研究进展［J］.第二军医大学报，2011，32(7)：790-794.
GAO Y，HUI N，LIU S R.Long non-coding RNA：research progress［J］.Academic Journal of Second Military Medical University，2011，32(7)：790-794.(in Chinese)
［6］MELLIN J R，COSSART P.The non-coding RNA world of the bacterial pathogen Listeria monocytogenes［J］.RNA Biol，2012，9(4)：372-378.
［7］HÄSLER R，JACOBS G，TILL A，et al.Microbial pattern recognition causes distinct functional micro-RNA signatures in primary human monocytes ［J］.PLoS One，2012，7(2)：e31151.
［8］MRAHEIL M A,BILLION A,KUENNE C,et al.Comparative genome-wide analysis of small RNAs of major Gram-positive pathogens：from identification to application ［J］. Microb Biotechnol，2010，3(6)：658-676.
［9］ZHANG Z C，MENG Q L，QIAO J，et al.RsbV of Listeria monocytogenes contributes to regulation of environmental stress and virulence［J］.Arch Microbiol，2013，195(2)：113-120.
［10］SIEVERS S，LILLEBAEK E M S，JACOBSEN K，et al.A multicopy sRNA of Listeria monocytogenes regulates expression of the virulence adhesin LapB［J］.Nucleic Acids Res，2014，42(14)：9383-9398.
［11］TOLEDO A A，DUSSURGET O，NIKITAS G，et al.The Listeria transcriptional landscape from saprophytism to virulence［J］.Nature，2009，459(7249)：950-956.
［12］乔军，孟庆玲，陈创夫，等.塔里木马鹿单核细胞增多性李氏杆菌的分离与鉴定［J］.中国农学通报，2008，24(1)：9-12.
QIAO J，MENG Q L，CHEN C F，et al.Isolation and identification of wild strain of Listeria monocytogenes in Tarim Wapiti［J］.Chinese Agricultural Science Bulletin，2008，24(1)：9-12.(in Chinese)
［13］GEORG J，HESS W R.Cis-antisense RNA，another level of gene regulation in bacteria［J］.Microbiol Mol Bio Rev，2011，75(2)：286-300.
［14］郭宏华，贾芙蓉，韩晓英，等.单核细胞增生性李斯特菌研究进展［J］.中国实验诊断学，2013，17(1)：197-199.
GUO H H，JIA F R，HAN X Y，et al.Advance in the research of Listeria monocytogenes［J］.Chinese Journal of Laboratory Diagnosis，2013，17(1)：197-199.(in Chinese)
［15］PEREZ J T，VARBLE A，SACHIDANANDAM R，et al.Influenza A virus-generated small RNAs regulate the switch from transcription to replication［J］.Proc Natl Acad Sci USA，2010，107(25)：11525-11530.
［16］RAASCH P，SCHMITZ U，PATENGE N，et al.Non-coding RNA detection methods combined to improve usability，reproducibility and precision［J］.BMC Bioinformastic，2010，11：491.
［17］EULALIO A，SCHULTE L，VOGEL J.The mammalian microRNA response to bacterial infections［J］.RNA Biol，2012，9(6)：742-750.
［18］孟庆玲，乔军，才学鹏，等.单核细胞增多性李氏杆菌野毒株 InlB 的分子特征及其表达和纯化研究［J］.微生物学报，2007，47(6)：1098-1101.
MENG Q L，QIAO J，CAI X P，et al.Molecular characteristics of InlB protein of Listeria monocytogenes wild strain and it’s expression and purification［J］.Acta Microbiologica Sinica，2007，47(6)：1098-1101.(in Chinese)
［19］张小娟，张荣光.影响电穿孔法转化效率的因素［J］.医学综述，2011，17(5)：666-668.
ZHANG X J，ZHANG R G.Factors which influence the efficiency of electroporation［J］.Medical Recapitulate，2011，17(5)：666-668.(in Chinese)

[1]	Lü Ruoyi, SI Xiaohui, SUN Zhigang, SHI Xiaomin, LIU Xiaoye. Drug Resistance Situation of Streptococcus suis and Prevention Measures of Infections [J]. Acta Veterinaria et Zootechnica Sinica, 2023, 54(12): 4920-4933.
[2]	ZHI Yan, MEI Chen, LIU Zhenyi, WUYUN Gerile, WANG Hongjun, HU Ge. Research Progress on the Virulence Factors of Avibacterium paragallinarum [J]. Acta Veterinaria et Zootechnica Sinica, 2023, 54(12): 4934-4942.
[3]	WANG Jianing, ZHANG Ziqiang, KONG Dejing, FENG Caicai, ZHANG Feike, LIU Yumei. Isolation and Identification of Klebsiella pneumoniae in Rabbits [J]. Acta Veterinaria et Zootechnica Sinica, 2023, 54(12): 5198-5206.
[4]	LI Lili, CHEN Kaifeng, CHEN Bing, ZHOU Zhouping, WANG Nanwei, QU Xiaoyun, XU Chenggang, LIAO Ming, ZHANG Jianmin. Regulatory Role of STM1827 in the Biofilm Formation and Environmental Stress of Salmonella Typhimurium [J]. Acta Veterinaria et Zootechnica Sinica, 2023, 54(12): 5207-5217.
[5]	YANG Menglin, ZHENG Shiqi, PENG Kai, WANG Wei, HUANG Yanhua, PENG Jie. Isolation and Identification of Pigeon-derived Salmonella Typhimurium and Pathogenic Analysis [J]. Acta Veterinaria et Zootechnica Sinica, 2023, 54(11): 4880-4888.
[6]	WU Zhouhui, WANG Yu, DU Heng, WANG Zhiwen, XIAO Shuang, WU Jinliang, WANG Zhen. Analysis of the Antibacterial Sensitization Activity of Tirapazamine against Multi-Drug Resistant Salmonella [J]. Acta Veterinaria et Zootechnica Sinica, 2023, 54(10): 4362-4371.
[7]	LIU Xinhuan, YUN Jialei, MAO Li, LI Jizong, HAO Fei, HE Miaofeng, YANG Leilei, ZHANG Wenwen, CHENG Zilong, SUN Min, LIU Maojun, WANG Shaohui, BAI Juan, LI Wenliang. Isolation, Identification, Virulence Genes and Drug Resistance Analysis of Escherichia coli Isolated from Diarrheal Goat and Sheep [J]. Acta Veterinaria et Zootechnica Sinica, 2023, 54(8): 3445-3454.
[8]	HU Xiuhua, SUN Zhixin, ZHAO Mengyang, XIE Jiaqi, WANG Min, CHEN Hailiang, GE Xin, LIU Tianlong, WANG Shaolin. Pathogenicity and Resistance Analysis of Enterococcus faecium from Wild Squirrels [J]. Acta Veterinaria et Zootechnica Sinica, 2023, 54(7): 3012-3021.
[9]	ZHAO Feifei, LI Jie, HAN Ning, XIE Shiting, ZENG Zhenling. Antibacterial Drug Resistance Analysis of Klebsiella pneumoniae Isolated from Slaughterhouse [J]. Acta Veterinaria et Zootechnica Sinica, 2023, 54(7): 3044-3053.
[10]	CHEN Songbiao, SHANG Ke, DU Fuxi, YU Zuhua, LI Jing, JIA Yanyan, LIAO Chengshui, ZHANG Chunjie, DING Ke, CHENG Xiangchao. Research Process of Assembly, Structural Features, and Secretion Regulatory Networks of Type VI Secretion System in Salmonella [J]. Acta Veterinaria et Zootechnica Sinica, 2023, 54(6): 2252-2263.
[11]	ZENG Chengrong, WANG Na, BI Wenwen, MEI Shihui, HE Guangxia, ZHANG Junjie, CHEN Ze, WEN Ming, ZHOU Bijun. Metabonomics Analysis of Duck Ileum Infected by Clostridium perfringens Type A [J]. Acta Veterinaria et Zootechnica Sinica, 2023, 54(6): 2555-2569.
[12]	LI Wei, ZHANG Qiang, QU Jiahao, WU Yaping, HU Ruochen, JIA Ruoyi, GUO Ruhai, MA Qingyi, PAN Guanglin, WANG Xinglong. Analysis on the Age Succession of Intestinal Flora of Giant Panda [J]. Acta Veterinaria et Zootechnica Sinica, 2023, 54(6): 2619-2630.
[13]	ZHAI Yunyi, YUAN Ye, LI Junmei, TIAN Lulu, DIAO Ziyang, LI Bin, CHEN Jialu, ZHOU Dong, JIN Yaping, WANG Aihua. Preparation and Preliminary Application of Monoclonal Antibody to Brucella Outer Membrane Protein 16 [J]. Acta Veterinaria et Zootechnica Sinica, 2023, 54(5): 2083-2091.
[14]	TIAN Yanhong, YU Jiangxu, JIAO Yuzhou, GAO Dongyang, CAI Xuwang. Research Progress on Structural Modification and Its Effects of Salmonella Lipopolysaccharides [J]. Acta Veterinaria et Zootechnica Sinica, 2023, 54(4): 1392-1402.
[15]	CHEN Yexin, XIE Mengyuan, LI Wenhao, ZHANG Zhidan, WANG Xiaodan, CHEN Kejia, LIU Pingping, ZHOU Weiguang, WANG Jianlong, XU Xiaojing. Analysis of Intestinal Flora in Faecal Samples from Rotavirus and Shigella Positive Calves with Diarrhoea [J]. Acta Veterinaria et Zootechnica Sinica, 2023, 54(4): 1624-1631.

Construction，Identification of ncRNA rli87 Gene Deletion Strains Listeria monocytogenes and Its Growth Characteristics

PDF (PC)

Knowledge

Abstract

Cite this article

share this article

References

Related Articles 15

Recommended Articles

Metrics

Comments