摘要
目的获得基因BC023882全长真核表达载体及检测该基因对培养细胞生长的影响。方法利用逆转录-多聚酶链反应(RT-PCR)扩增基因BC023882的编码区序列,克隆到pcDNA3.1(-)/Myc-H is(+)/lac Z真核表达载体中,构建基因BC023882的真核表达载体;脂质体法转染小鼠胚胎成纤维细胞和COS-7细胞,并且通过细胞生长曲线测定和流式细胞仪观察该基因对细胞生长的影响。结果酶切并测序鉴定证明获得的重组子符合基因BC023882的编码序列;转染该基因后细胞增殖减慢,细胞增殖指数减低。结论成功构建了基因BC023882的真核表达载体,并且观察到该基因对细胞生长起到一定的抑制作用。
Objective To obtain eukaryotic expression vectors containing coding region of the gene BC203882 and detect its effect on cell growth. Methods The coding region of the gene BC023882 was amplified by RT-PCR, then cloned into eukaryotic expression vector pcDNA3.1 （ - ）/Myc-His （ ＋ ）/lacZ. The recombinant plasmid was transfected into embryonic fibroblast of mouse and COS-7 cells with liposome method. Its effects on cell proliferation were examined by FACS and growth curves. Results The eukaryotic expression vectors containing coding region of the gene BC203882 were constructed. The growth of the transfected cells was retarded, and the proliferation index of cells was significantly decreased. Conclusion The eukaryotic expression vectors of the gene BC023882 were eonstrueted sueeessfully and the inhibitory effect of the gene on eell growth was observed.
出处
《第三军医大学学报》
CAS
CSCD
北大核心
2006年第11期1178-1180,共3页
Journal of Third Military Medical University
基金
全军医学科学技术研究"十五"计划基金重点课题(01Z068)~~
