摘要
目的 :提取人胎盘组织中RNA ,以其中的mRNA为模板 ,扩增人 βNGFcDNA ,并纯化 ,为进一步克隆及表达 βNGFcDNA奠定基础。 方法 :异硫氢酸胍法提取RNA ;采用RT PCR技术扩增人 βNGFcDNA ;柱离心方法纯化。结果 :从人胎盘组织中提取出RNA ,以其中的mRNA为模板 ,扩增出人 βNGFcDNA ,经琼脂糖凝胶电泳鉴定为目的片段 ,并回收纯化。结论 :以人胎盘组织中的mRNA为模板 ,运用RT PCR技术扩增βNGFcDNA ,同时在 5′端以限制性内切酶酶切位点加以修饰 。
Objective: RNA was extracted from human placenta. Human β nerve growth factor complementary DNA(β NGF cDNA) was amplified by using mRNA as template and then purified. All of these works are the basis of further study of cloning and expression of β NGF cDNA. Methods: RNA was extracted by guanidine isothiocyanate methodHuman βNGF cDNA was amplified by RT-PCR technologyThe βNGF cDNA was purified by column centrifugalization. Results: RNA was extracted from the tissue of human placentaHuman βNGF cDNA was amplified, mRNA of RNA served as template .The segment was identified as target DNA by agarose electrophoresis ,and then purified.Conclusion: Human βNGF cDNA was amplified by RT-PCR technology using mRNA from human placenta as template and cDNA 5′end was modified with cutting-site of restriction endonuclease, which is convenient for the cloning and expression of βNGF gene.
出处
《中国医科大学学报》
CAS
CSCD
北大核心
2001年第3期169-170,173,共3页
Journal of China Medical University
