唾液富组蛋白5克隆载体的构建  

Constructing the Cloning Vector of Salivary Histatin 5

作　　者：金科华[1] 周琦[2] 罗德生[1] 王彪[2] 程新潮[2] 段海瑞[2] 

机构地区：[1]咸宁学院医学院生物化学教研室,湖北咸宁437100 [2]咸宁学院医学院

出　　处：《咸宁学院学报（医学版）》2007年第5期385-387,共3页Journal of Xianning Univarsity(medical Sciences)

摘　　要：目的将唾液富组蛋白5(histatin5)的cDNA克隆至T载体,获取大量酶切后的目的片段。方法选用大肠杆菌偏爱密码子编码histatin 5氨基酸,设计5′端带酶切位点、3′端互补的两条引物,通过overlap PCR获取histatin 5的cDNA,将cDNA与T载体连接后转化大肠杆菌,筛选阳性克隆,PCR、酶切、测序鉴定。酶切重组T载体,获取目的片段。结果Histatin 5的cDNA正确克隆至T载体,无突变。结论克隆载体构建成功,有利于目的片段的酶切和回收。Objective To construct a cloning vector containing the cDNA of salivary histatin 5, acquire large amount of digested cDNA. Methods Histatin 5 was encoded by opitirnal codon of E. coli, a pair of comple- mentary primers were designed and used to synthsize the cDNA of histatin 5 by overlap PCR, the cDNA was flanked with EcoRIand Sail sites. The cDNA was linked with T vector and transfered to E. coli, the positive recombinants were screened and identified by PCR, digestion and sequencing. Digested cDNA was obtained by digesting the positive recombinants. Results The cDNA of histatin 5 was correctly inserted into T vector wihout mutation. Condusion Cloning vector was successfully constructed, digested cDNA could be harvested more easily.

关 键 词：唾液富组蛋白5 克隆 T载体 

分 类 号：Q786[生物学—分子生物学]