肝脏再生增强因子cDNA的克隆及稳定细胞株的建立  

Clone of the human augmenter of liver regeneration cDNA and establishment of steady cells

作　　者：曾晓波[1] 苏先狮[1] 

机构地区：[1]中南大学湘雅二医院肝病研究所,湖南省长沙410011

出　　处：《中国基层医药》2007年第9期1447-1448,1585,共3页Chinese Journal of Primary Medicine and Pharmacy

摘　　要：目的通过对人源肝脏再生增强因子(ALR)cDNA进行克隆及活性的初步鉴定,为基因工程制备重组人肝脏ALR(hALR)提供资料。方法提取人胎肝总RNA,通过RT-PCR获得hALR cDNA,构建真核表达质粒pcDNA3.1-ALR,转化大肠杆菌扩增质粒。将质粒pcDNA3.1与pcDNA3.1-ALR分别转染HepG2细胞,G418筛选建立稳定细胞株,MTT法检测hALR的生物学活性。结果测序证实克隆得到的hALR序列完全正确。MTT比色结果,转染hALR的细胞增殖高于转染空质粒和未转染细胞(P<0.05)。结论hALR具有促肝细胞增殖的作用。Objective To provide basic feature of preparation recombinant hALR by gene engineering in larger scale. Methods The human ALR cDNA was obtained by using RT-PCR method with total RNA extracted from the fetal hepatic tissue,then was cloned into the vector pcDNA3.1 and transformed into E. coli BL21 for enlarging vector pcDNA3.1-hALR. The activity of hALR was evaluated by MTT assay. Results The plasmid pcD- NA3.1-hALR was proved to be correct by sequencing. MTT assay showed the index＇of proliferation of pcDNA3.1- ALR-HepG2 was significantly higher than pcDNA3.1-HepG2 and HepO2 of the control group （P 〈 0.05）. Conclusion The activity showed that hALR has potential effects in prompting the liver cell regeneration.

关 键 词：肝脏再生增强因子 克隆 稳定细胞株 

分 类 号：R346[医药卫生—基础医学]