鸡输卵管组织特异性分泌表达载体的构建和表达 |
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引用本文: | 孙明军,赵晨,沙金,韩海棠,郑克俭,李赞东.鸡输卵管组织特异性分泌表达载体的构建和表达[J].农业生物技术学报,2003,11(1):75-78. |
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作者姓名: | 孙明军 赵晨 沙金 韩海棠 郑克俭 李赞东 |
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作者单位: | 中国农业大学生物学院,北京,100094 |
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基金项目: | 国家自然科学基金资助(39980030). |
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摘 要: | 摘要:为了实现外源基因在鸡(Gallus gallus )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450 bp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1.2 kb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。
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关 键 词: | 关键词:卵清蛋白5ˊ调控序列 分泌表达载体 溶菌酶 绿色荧光蛋白 |
修稿时间: | 2002年1月23日 |
Construction and Expression of the Specific Secretory Vector in the Chicken Oviduct Cells |
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Abstract: | Abstract: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. |
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