鸡输卵管组织特异性分泌表达载体的构建和表达  

Construction and Expression of the Specific Secretory Vector in the Chicken Oviduct Cells

作　　者：孙明军[1] 赵晨[1] 沙金[1] 韩海棠[1] 郑克俭[1] 李赞东[1] 

机构地区：[1]中国农业大学生物学院,北京100094

出　　处：《农业生物技术学报》2003年第1期75-78,共4页Journal of Agricultural Biotechnology

基　　金：国家自然科学基金资助(39980030)。

摘　　要：为了实现外源基因在鸡(Gallusgallus )输卵管上皮细胞内的特异性分泌表达,采用RT-PCR的方法扩增了鸡溶菌酶基因的450bp的cDNA片段,该片段保留了5ˊ端信号肽编码序列。将该片段插入到含有1.2kb卵清蛋白5ˊ调控序列的特异性表达载体pOVA-GFP中,与绿色荧光蛋白基因形成融合基因,使融合蛋白具有分泌功能。新构建的分泌表达载体pOVALG转染鸡胚成纤维细胞和大鼠乳腺上皮细胞系SHZ-88后,成功地检测到了绿色荧光蛋白的表达。To express foreign proteins in chicken (Gallus gallus ) oviduct cells and secret to oviduct, 450 bp chicken lysozyme cDNA fragment was cloned by using RT-PCR method. Treated by the restriction endonuclease Bam HⅠand Hind Ⅲ, the fragment was ligated with the vector generated by the same Bam HⅠ-HindⅢ digestion of plasmid pOVA-GFP. The resulting expressive vector pOVALG included an 1.2 kb chicken ovalbumin 5'-flanking regulatory region and a lysozyme-GFP fusion gene with the signal peptide of the lysozyme. After transfecting the vector to the chicken embryo fibroblasts and mammary cancer cell line of rat SHZ-88 cells, green fluorescent protein had been detected successfully.

关 键 词：卵清蛋白5 调控序列 分泌表达载体 溶菌酶 绿色荧光蛋白 

分 类 号：Q786[生物学—分子生物学]