人β-神经生长因子基因重组真核表达载体的构建  

Construction of recombinant eukaryotic expression vector carrying human β-NGF gene

作　　者：陈维忠[1] 韩晓枫[2] 郝爱民[2] 张澜生[3] 

机构地区：[1]江苏省医学会,南京210008 [2]无锡市第二人民医院检验科 [3]苏州医学院细胞免疫研究中心

出　　处：《临床检验杂志》2000年第5期261-263,共3页Chinese Journal of Clinical Laboratory Science

基　　金：江苏省自然科学基金!(编号 :BS98310 )资助

摘　　要：目的　构建 β 神经生长因子 (β NGF)基因重组真核表达载体 ,为 β NGF基因治疗和临床实验诊断研究打下良好的基础。方法　将质粒PGEM β NGF进行酶切获得 β NGF基因片段 ,用T4连接酶将其插入真核表达载体PcDNA3;以T7、P2为引物进行PCR ,论证插入片段的方向 ;经测序和酶切对插入片段进行分析和进一步鉴定。结果　PCR产物琼脂糖电泳结果显示 :在预期位置有阳性条带 ;序列分析和酶切结果证实插入片段序列正确。结论　β NGF基因重组真核表达载体PcDNA3 β NGF构建成功 ,为进一步开展 β NGF基因治疗神经系统疾病和临床实验诊断奠定了基础。Objective To construct recombinant eukaryotic expression vector of human β*.NGF to provide the scientific basis of research of gene therapy and clinical experiment diagnosis.Methods Human β*.NGF cDNA obtained from PGEM*.β*.NGF by enzyme digestion was inserted into an eukaryotic expression vector,PcDNA 3 with T4 ligase.The recombinant plasmid PcDNA 3*.β*.NGF was verified by PCR with T7 and P2 primer,squencing and enzyme digestive analysis.Result A positive band was formed when product of PCR electrophresed on agar gel and inserted fragment was confirmed correctly by sequencing and enzyme digestion analysis.Conclution The recombinant eukaryotic expression vector,PcDNA 3*.β*.NGF was constructed successfully for the gene therapy of the nervous system diseases and clinical experiment diagnosis.

关 键 词：Β-神经生长因子 重组载体 序列分析 基因治疗 

分 类 号：Q516[生物学—生物化学] R394[医药卫生—医学遗传学]