论文部分内容阅读
目的 :克隆乳清酸蛋白基因 (WAP)启动子并构建其载体。方法 :以PCR技术克隆WAP启动子 2 .4kbDNA片段和加polyA信号 0 .37kbDNA片段 ;应用体外连接技术 ,连接pBluescript质粒、WAP启动子和加polyA信号DNA片段 ;转化连接产物 ,以内切酶酶切、琼脂糖电泳鉴定阳性克隆 ,对插入片段进行序列分析。结果 :琼脂糖电泳显示 ,WAP启动子和加polyA信号DNA片段克隆成功 ;内切酶酶切鉴定阳性克隆 ,酶切片段与预期长度一致。插入片段序列分析与文献报道一致。结论 :WAP启动子克隆及其乳腺特异性表达载体pWA构建成功
OBJECTIVE: To clone the whey acid protein (WAP) promoter and construct its vector. Methods: The 2.4 kb DNA fragment of WAP promoter and 0. 37 kb DNA fragment with polyA signal were cloned by PCR. The pBluescript plasmid, WAP promoter and polyA signal DNA fragment were ligated by using in vitro ligation technology. , Positive clones were identified by agarose gel electrophoresis, and the inserted fragments were sequenced. Results: The agarose gel electrophoresis showed that the WAP promoter and polyA signal DNA fragment were successfully cloned. The positive clones were identified by restriction endonuclease digestion and the fragments were consistent with the expected length. Sequence analysis of inserted fragments was consistent with that reported in the literature. Conclusion: The WAP promoter cloning and mammary gland-specific expression vector pWA were successfully constructed