长白猪甘露聚糖结合凝集素A基因的 克隆与原核表达

中国农学通报 ›› 2008, Vol. 24 ›› Issue (9): 4-8.

所属专题：生物技术；畜牧兽医

长白猪甘露聚糖结合凝集素A基因的
克隆与原核表达

姚欣,刘玉芬,唐高霞,刘洪雨

收稿日期:2008-06-24 修回日期:2008-07-28 出版日期:2008-09-05 发布日期:2008-09-05

Cloning and Prokaryotic Expression of Porcine Mannan-binding Lectin A Gene

Yao Xin, Liu Yufen, Tang Gaoxia, Liu Hongyu

Received:2008-06-24 Revised:2008-07-28 Online:2008-09-05 Published:2008-09-05

摘要/Abstract

摘要： 从长白猪肝脏细胞中提取总RNA，通过RT-PCR法扩增pMBL-A基因，与pMD18-T载体连接进行TA克隆，得到猪甘露聚糖结合凝集素A（porcine mannan-binding lectin A，pMBL-A）基因。再亚克隆到pPROEXHTb表达载体上，构建重组表达质粒。将阳性重组表达质粒转化到大肠杆菌中诱导表达重组pMBL-A蛋白，通过SDS-PAGE检测蛋白的表达情况。结果成功扩增得到包括完整开放读码框长为875bp的全长cDNA片段，并原核表达了重组蛋白，为进一步研究该蛋白的遗传特征以及为猪遗传育种方面的研究提供依据。

关键词: 固始鸡, 固始鸡, 孵化, 钙, 磷

Abstract: Total RNA was extracted from liver cells of landrace, pMBL-A gene was amplified by RT-PCR. It was cloned to pMD18-T vector, then the mannan-binding lectin A gene of landrace was obtained. It was subcloned to pPROEXHTb vector for expression, recombinant expression plasmid was constructed. The recombinant expression plasmid was transformed into E.coli and was induced to express recombinant protein of pMBL-A. The expressed protein was identified by SDS-PAGE. The cDNA of pMBL-A gene was successfully cloned, its full-length was 875bp including the whole ORF. Recombination protein was pronuleus expressed, it provides affordance for the research of genetic character of the gene and genetic breeding of pigs.

中图分类号:

姚欣,刘玉芬,唐高霞,刘洪雨. 长白猪甘露聚糖结合凝集素A基因的
克隆与原核表达[J]. 中国农学通报, 2008, 24(9): 4-8.

Yao Xin, Liu Yufen, Tang Gaoxia, Liu Hongyu. Cloning and Prokaryotic Expression of Porcine Mannan-binding Lectin A Gene[J]. Chinese Agricultural Science Bulletin, 2008, 24(9): 4-8.

参考文献

[1] Jack DL,Turner MW.Anti-microbial activities of mannose-binding lectin. Biochem Soc Trans,2003,31:753-757.
[2] Cedzynski M,Madalinski K,Gregorek H,et al.Possible disease-modifying factors: the mannan-binding lectin pathway and infections in hereditary angioedema of children and adults.Arch Immunol Ther Exp (Warsz),2008,56(1):69-75.
[3] Phaneuf LR,Lillie BN,Hayes MA,et al.Binding of mouse mannan-binding lectins to different bacterial pathogens of mice.Vet Immunol Immunopathol, 2007,118(1-2):129-133.
[4] Phatsara C,Jennen DG,Ponsuksili S,et al.Molecular genetic analysis of porcine mannose-binding lectin genes,MBL1 and MBL2,and their association with complement activity.Int J Immunogenet,2007,34(1):55-63.
[5] Butler GS,Sim D,Tam E,et al.Mannose-binding lectin(MBL) mutants are susceptible to matrix metalloproteinase proteolysis[J].J Biol Chem,2002, 277(20):17511-17519.
[6] Turner M.W. The role of mannose-binding lectin in health and disease.Molecular Immunology,2003,40:423-429.
[7] Thielens NM,Tacnet-Delorme P, Arlaud GJ.Interaction of C1q and mannan-binding lectin with viruses.Immunobiology,2002,205:563-574.
[8] van de Wetering JK,van Golde LM,Batenburg JJ.Collectins:players of the innate immune system.Eur J Biochem,2004,271:1229-1249.
[9] Holmskov U,Thiel S,Jensenius JC.Collectins and ficolins:humoral lectins of the innate immune defense.Annu Rev Immunol,2003,21:547-578.
[10] Dahl IR,Thiel S,Matsushita M, et al. MASP-3 and its association with distinct complexes of the mannan-binding lectin complement activation pathway. Immunity,2001,15:127-135.
[11] Agah A,Montalto MC,Young K,et al.Isolation,cloning and functional characterization of porcine mannose-binding lectin. Immunology,2001, 102:338-343.
[12] Brandon N.Lillie,Natalie D.Keirstead,E.James Squires,et al.Single-nucleotide polymorphisms in porcine mannan-binding lectin A.Immunogenetics, 2006,58:983-993.
[13] Presanis JS,Kojima M,Sim RB. Biochemistry and genetics of mannan-binding lectin(MBL).Biochem Soc Trans,2003,31:748-752.
[14] Summerfield JA.Clinical potential of mannose-binding lectin replacement therapy.Biochem Soc Trans,2003,31(4):770-773.
[15] Thomas Vorup-Jensen,Esben S,Sorensen,et al.Recombinant expression of human mannan-binding lectin.Int Immunopharmaco,2001,1:677-687.
[16] Jensenius JC,Jensen PH,Mc Guire K,et al.Recombinant mannan-binding lectin(MBL) for therapy.Biochem Soc Trans,2003,31:763-767.
[17] Vorup-Jensen T,Sorensen ES,Jensen UB,et al.Recombinant expression of human mannan-binding lectin[J].Int Immunopharmacol,2001,1(4):677-687.

长白猪甘露聚糖结合凝集素A基因的
克隆与原核表达

Cloning and Prokaryotic Expression of Porcine Mannan-binding Lectin A Gene

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长白猪甘露聚糖结合凝集素A基因的 克隆与原核表达

Cloning and Prokaryotic Expression of Porcine Mannan-binding Lectin A Gene

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长白猪甘露聚糖结合凝集素A基因的
克隆与原核表达