嗜热菌β-糖苷酶基因乳腺特异性表达载体鉴定

IDENTIFICATION OF MAMMARY GLAND SPECIFIC EXPRESSION VECTOR FOR Ssβ-GLY

本研究目的在于对实验室优化构建的嗜热菌β-糖苷酶基因乳腺特异性表达载体PLOX-LacS的功能进行鉴定,检测目的基因能否正常表达。实验通过脂质体介导方法将载体质粒转染小鼠的乳腺癌细胞,筛选获得的阳性细胞通过PCR和RT-PCR的方法进行嗜热菌β-糖苷酶基因的整合检测和表达检测。基因整合PCR鉴定结果表明小鼠乳腺癌细胞基因组中整合有嗜热菌糖苷酶基因,目的片段大小为1203bp;RT-PCR表达鉴定结果显示,目的片段大小与预期相同,说明诱导后的小鼠乳腺癌细胞表达了完整的嗜热菌β-糖苷酶基因。实验表明该载体能够使目的基因整合在乳腺细胞基因组中并特异性表达。为后期生产转基因奶牛提供实验材料。

The purpose of this study is to identify the function of mammary gland specific expression vector （ PLOX - LacS） for Ss - gly, and detect whether the target genes normally expressed. Verification of expression vector by mammary gland cancer cell through the method of PCR and RT - PCR analysis. The gene integration detection by PCR showed that mouse breast cancer cell genome integrated the LacS gene target fragment size 1203bp ; RT - PCR expression identification results show that the mouse breast cancer cells which are induced expression the LacS gene target fragment. Conclusion： The experiments show that the vector can intergrate with target gene and espmssed specifically in mammary cells genome . Provide experimental material for the late production of transgenic cows .