摘要
目的 构建含鼠连接蛋白 4 3基因的真核表达重组质粒。 方法 从胎鼠中提取总 RNA,经 RT- PCR方法获得并扩增含全部编码序列的连接蛋白 4 3c DNA片段 ,将其克隆到 p Bud CE4 .1真核表达载体上 ,并进行酶切鉴定和测序分析。 结果 RT- PCR技术扩增出约 1 .1 kb的连接蛋白 4 3基因全部编码序列的片段。测序分析证实所插入连接蛋白 4 3基因片段的序列完全正确。 结论 鼠连接蛋白 4
Objective To construct eukaryotic expression recombinant plasmid containing mouse connexin 43 gene. Methods Total RNA was extracted from embryo mice. Connexin 43 cDNA was amplified by RT PCR, isolated and inserted into pBudCE4 1 eukaryotic expression plasmid. Then the recombinant plasmid was identified by enzyme digestion analysis and sequencing. Results The connexin 43 gene fragments of about 1 1 kb were specifically amplified by RT PCR technique. The recombinant plasmid digested by Hind Ⅲ and BamH Ⅰ was coincident with the expected connexin 43 cDNA fragment. The gene fragment containing of the whole coding sequence was successfully cloned in pBudCE4 1 plasmid. Conclusion Mouse recombinant plasmid pBudCE4 1/connexin 43 was successfully constructed.
出处
《福建医科大学学报》
2004年第3期271-274,共4页
Journal of Fujian Medical University
