摘要
目的构建人RhoC基因正、反义真核表达载体,研究RhoC基因对胆管癌QBC939细胞株增殖的影响。方法应用分子克隆技术,用KpnⅠ和BamHⅠ双限制性内切酶从RhoC基因切下所需目的片段,然后将该片段正、反向定向克隆到真核表达载体pcDNA 3.1/Hyg(+)上,构建正、反义RhoC cDNA真核表达载体,以脂质体转染人胆管癌QBC939细胞,检测细胞生长曲线。结果经测序鉴定,将正、反义目的片段成功的连接到pcDNA 3.1/Hyg(+)载体上。结果序列测定证实RhoC基因正、反义真核表达载体成功构建,反义RhoC基因抑制QBC939细胞增殖,正义RhoC基因促进QBC939细胞增殖。结论成功地将RhoC目的片段正、反向插入到pcDNA 3.1/Hyg(+)中,构建了人RhoC正、反义表达真核载体,RhoC基因调控QBC939细胞增殖。
Objective To construct the sense and antisense expression vector of human RhoC and observe the relationship between RhoC gene and tumor proliferation. Methods RhoC gene fragment, cleaved from cDNA RhoC with Kpn T and BamH T was cloned into the pcDNA3.1/Hyg（＋） plasmid to reconstruct sense and antisense eukaryotic expression vector. The constructed recombinant was identified by agaro6e gel electrophoresis and sequenced. Results DNA sequence analysis and agarose gel electrophoresis confirmed that the sense and antisense target fragment was successfully bound to pcDNA3.1/ Hyg （＋）. Antisense RhoC gene inhibited proliferation of QBC939, while sense RhoC gene promoted proliferation of QBC939. Conclusion The sense and antisense eukaryotic expression vector of RhoC has been constructed successfully by means of inserting the sense and antisense target,fragment into pcDNA3. 1/Hyg （＋）. RhoC gene could control proliferation of QBC939.
出处
《中华实验外科杂志》
CAS
CSCD
北大核心
2005年第11期1289-1291,共3页
Chinese Journal of Experimental Surgery
