人IL-3、GM-CSF和Hly基因的cDNA克隆及测序  

Cloning and Sequencing of Human Genes cDNA of the IL-3、GM-CSF and Lysozyme

作　　者：孙英民[1] 孙朕[1] 樊宝良[1] 

机构地区：[1]河北农业大学动物科技学院,河北保定071000

出　　处：《中国畜牧兽医》2011年第4期91-94,共4页China Animal Husbandry & Veterinary Medicine

基　　金：农业部转基因重大专项项目"高品质转基因奶牛新品种培育"(2008ZX08007-001);农业部转基因重大专项项目"环境友好的转基因猪新品系培育"(2008ZX08006004);国家863计划重点项目"家禽与昆虫蛹生物反应器研究"(2007AA100504)

摘　　要：试验利用PCR技术扩增并获得了人IL-3、GM-CSF和人溶菌酶(human lysozyme,Hly)基因组基因编码区序列,人IL-3和GM-CSF基因PCR产物与pMD19-T连接,Hly基因PCR产物同pEASY-Blunt载体连接,转化大肠杆菌JM109感受态细胞,蓝白斑筛选后经PCR及酶切鉴定,选取测序正确的克隆连接到pIRESneo载体中构建相关真核表达载体,分别用这些基因的表达载体转染PK15细胞,48 h后提取转染细胞的总RNA,并采用RT-PCR方法获得了人IL-3、GM-CSF和Hly基因的cDNA,结果发现,所获得的cDNA序列同NCBI登录的序列完全同源。The coding region of human genomic genes of the IL-3,GM-CSF and lysozyme（Hly） were obtained by PCR method.The IL-3 and GM-CSF genes were linked with the pMD19-T vector,and the Hly gene was linked with the pEASY-Blunt vector.Then the eukaryotes expression vector for these genes were constructed by insert these gene fragments into pIRESneo vector.Following,transfection the PK15 cells with these eukaryotes expression vector respectively.The total RNA of the transfected cells were extracted after 48 hours and the cDNA of the human genes of the IL-3,GM-CSF and Hly were obtained by RT-PCR method.The results showed that the sequences were completely homologous with the registered ones of the NCBI.

关 键 词：IL-3基因 GM-CSF基因 Hly基因 CDNA 克隆 测序 

分 类 号：Q785[生物学—分子生物学]