摘要
构建Hlrg基因的定点突变体,研究突变Hlrg基因表达的蛋白对HepG2细胞的影响。首先利用数据库对Hlrg基因的结构特点进行分析,在此基础上用PCR法构建Hlrg基因的定点突变体。将突变的Hlrg基因克隆到pcDNA3.1(+)载体中,并稳定转染到HepG2细胞中。流式细胞仪和透射电镜观察突变基因表达的蛋白对HepG2细胞的影响。结果获得了针对Hlrg基因的定点突变体,亮氨酸拉链中的第二个亮氨酸被突变成丝氨酸。发现稳定表达HLrgm蛋白的HepG2细胞中出现一定比例的凋亡细胞。表明构建了Hlrg基因定点突变体,为进一步的功能研究奠定了基础。
To construct site-directed mutants of human lrg and to observe the influence of HepG2 by mutants of human Lrg, the structural character of human lrg was analyzed by computer, then Hlrg mutant was constructed by PCR method. Construction eukaryotic expressed vector pcDNA3.1 （ ＋ ） -Hlrgm, then transfected into HepG2 cells.The biology influence was surveyed by FCM and TEM. Site-directed mutant of human lrg were constructed successfully, the second leucine was switched to serine in the Leucine zipper domain. HepG2 Cells of stable over-expression Hlrgm were obtained. Compared to the control, apoptosis cell was displayed in HepG2 cells of stable over-expression Hlrgm. It is conclused that site-directed mutants of human lrg was constructed. These results may shed light on further study of human lrg.
出处
《科学技术与工程》
2006年第10期1341-1344,共4页
Science Technology and Engineering
基金
国家自然科学基金(30170361
30400413
30571586)
第四军医大学博士后基金(00001149)资助
