鼠防御素Crp4基因的克隆及序列分析  

The cDNA Cloning and Sequence Analysis of Defensin Crp4 from Kunming Mouse Ileum

作　　者：陈宝玉[1] 查云峰 徐兴然[3] 张慧丽[4] 邓旭明[1] 

机构地区：[1]吉林大学畜牧兽医学院,长春130062 [2]广东省动物防疫监督总所,广州510230 [3]西南农业大学动物科技学院,重庆400617 [4]吉林大学植物科技学院,长春130062

出　　处：《吉林农业大学学报》2006年第1期88-92,共5页Journal of Jilin Agricultural University

基　　金：国家"924"项目资助(2004AA002060)

摘　　要：从昆明鼠回肠末端提取总RNA,经逆转录PCR(RT-PCR)扩增出鼠防御素Crp4基因片段。将回收的基因片段插入pMD-18T载体,小提质粒经限制性酶切鉴定后进行测序。结果表明:扩增出的基因片段长度为279 bp。该基因编码93个氨基酸,其中成熟肽由34个氨基酸组成。通过序列分析发现1对引物克隆到2个不同的防御素基因(Crp4-1和Crp4-2)中,与2个鼠防御素基因(AY746426和AY746425)的同源性分别为97.8%和97.1%。在与家鼠、人、猕猴和兔的防御素成熟肽序列进行比较后,发现它们均含有6个保守的半胱氨酸。这一发现有助于对鼠防御素的结构和功能进行深入研究。Total RNA from Kunming mouse ileval distal was isolated. The cDNA of mouse defensin was amplified by the reverse transcription polymerase chain reaction （RT-PCR）. After being inserted into cloning vector（pMD-18Tvector）, the obtained cDNA fragment was identified by restriction endonuclease and then sequenced. Its length is 279bp and encodes 93 amino acids, including mature sequence peptide whose length is 34 amino acids. Through sequence analysis, two different sequences （Crp4-1 and Crp4-2） obtained with one pair of primers were found, and their homogeneity is up to 97.8% and 97.1% respectively among Crp4-1, Crp4-2 and other two mouse defensins reported. Their comparison with house mouse, Homo sapiens, Macaca mulatto and rabbit defensin showed that their mature peptide contained the α-defensin consensus sequence of six invariantly spaced cysteine residues. The findings facilitate an advanced research of the function and structure in mouse defensin.

关 键 词：防御素 Crp4 克隆 序列分析 

分 类 号：R978.1[医药卫生—药品]