人角质细胞生长因子的植物表达载体构建

Construction of plant expression vector of human KGF

人角质细胞生长因子(KGF)有3个外显子,2个内含子．以人 DNA 为模板,PCR 扩增获得角质细胞生长因子外显子2和外显子3；合成单链的寡核苷酸,经链延伸和 PCR 扩增从而获得外显子1．应用延伸 PCR 将3个外显子连接得 KGF 编码序列,并连接 CaMV35S 启动子和 NOS 终止子后克隆至植物表达质粒 pBI 1301．

Human KGF has three extrons and two introns and in order to express KGF protein in the plant cell, we constructed the binary expressing vector. Firstly we synthesized the polynucleotide of extron 1 and obtained extron 1 of human KGF by the prolongation and PCR from human lung tissue; secondly we amplified extron 2 and extron 3 of KGF and conjoined three extrons by PCR. Thirdly CAMV35s-KGF-NOS was obtained after ligating KGF with CaMV35S and NOS. Finally pBI 1301-KGF was constructed after inserting CAMV35s-KGF-NOS into pBI 1301.