犬IGF-I编码区cDNA克隆及其序列分析

Cloning and sequencing of encoding gene of dog IGF-I

目的 :克隆犬胰岛素样生长因子 (IGF -I)编码区基因 ,构建重组真核表达载体。方法 :从犬左心室组织中提取总RNA ,通过RT -PCR获取IGF -1cDNA编码区全序列 ,将其与 pcDNA3 .1(+ )质粒载体连接 ,构建重组真核表达载体pcDNA3 .1(+ ) /IGF -1,转化大肠杆菌DH5α后 ,随机挑选数个克隆 ,提取质粒DNA ,通过限制性酶切和核苷酸序列鉴定阳性克隆。结果 :重组质粒pcDNA3 .1(+ ) /IGF -1的酶切图谱和序列分析结果与国外文献报道一致。结论 :克隆到IGF -1编码区基因 ,成功构建重组真核表达载体 pcDNA3 .1(+ ) /IGF -1。

AIM:To get the encoding gene of dog insulin -l ike growth factor I (IGFI). METHODS:Total RNA was extracted f rom dog left ventricular myocardium.The desired DNA product was obtained from th e total RNA by RT-PCR.The segment (about 297 bp) was inserted into pcDNA3.1(+) v ector and the inserted plasmid was transformed into E.coli DH5α.The positiv e clone was analyzed by restriction endonuclease mapping and DNA sequencing.RESULTS:The restriction endonuclease map and sequence of dog IGF- I functional fragment were consistent with those of the published.CONCL USION:The encoding gene of dog IGF-I has been cloned.