摘要
目的 :克隆人KGF_2基因 ,并在大肠杆菌中进行表达。方法 :通过PCR扩增 ,从人胎肝cDNA文库中获得编码人KGF_2的cDNA ,并将其分别克隆入pBV2 2 0 ,pLEX和pGEX4T_1质粒中 ,研究其在不同表达质粒和大肠杆菌中的表达情况。结果 :克隆得到的KGF_2cDNA经测序证明与文献报道的序列一致。将其克隆于pBV2 2 0质粒中 ,在E .coliJM10 9中表达量相对最高 ,约占菌体总蛋白的 4 % ;克隆于pLEX质粒中 ,在E .coliGI72 4中的表达量约占全菌总蛋白的 5 % ;克隆于pGEX4T_1中 ,在E .coliJM10 9中的表达量可占到全菌总蛋白的 2 0 %。结论 :采用GST融合蛋白表达质粒pGEX4T_1对KGF_2进行表达 ,较pBV2 2 0和pLEX具有明显优势 ,能实现对KGF_2的高效表达 ,为进一步的功能研究奠定了基础。
Objective:To clone the gene of human keratinocyte growth factor-2 (KGF-2) and express it in E.coli. Methods: KGF-2 cDNA was obtained from human fetal liver cDNA library by PCR and cloned into several plasmids respectively to investigate its expression in different E.coli strains. Results: Sequencing analysis showed that the obtained KGF-2 cDNA was consistent with that reported. When cloned into pBV220,the highest expression of KGF-2 was achieved in E.coli JM109, accounting for about 4% of the total cell protein. Cloned into pLEX and pGEX4T-1 respectively, the KGF-2 amounted to about 5% of total cell protein in former and 20% in latter. Conclusion: The results suggest that the gene of KGF-2 reaches a high level of expression by cloning into pGEX4T-1. It provides a reliable foundation for the functional investigation of KGF-2.
出处
《军事医学科学院院刊》
CSCD
北大核心
2004年第1期25-27,67,共4页
Bulletin of the Academy of Military Medical Sciences
