科技创新与应用2024，Vol.14Issue(16)：7-11,5.DOI:10.19981/j.CN23-1581/G3.2024.16.002
IGF-1基因克隆及其产物的测序鉴定
摘要
Abstract
Gene overexpression is a common method in basic experiments and gene therapy applications,and plasmid amplification is an indispensable technique in the process of gene overexpression.How to amplify and extract plasmids standardized and efficiently is the premise of gene therapy application,and it is also a difficult problem for many basic researchers.To establish the optimal scheme of amplification and extraction of overexpression plasmid on the basis of reducing the experimental cost by using the existing experimental technology.Firstly,the LB medium and competent cells were made by traditional method,and the transformed plasmid was amplified and cultured,and then the plasmid was extracted by commercial kit.After repeated experiments,the plasmid extraction process of commercial kit was optimized,the efficiency was improved,and the plasmid solution with high purity and concentration was obtained.The sequence of the target gene was verified by ultramicro spectrophotometer,agarose gel electrophoresis and sanger sequencing,which was consistent with that in NCBI database,indicating that the amplification and extraction were successful.The optimized plasmid amplification and extraction scheme is standard,efficient and low cost,which is the best choice for plasmid gene cloning.关键词
IGF-1基因/质粒克隆/基因过表达/质粒的扩增/基因治疗应用Key words
IGF-1 gene/plasmid cloning/gene overexpression/plasmid amplification/gene therapy application分类
生物科学引用本文复制引用
曹尚美,邹真真,陈泊霖,杨少哲,张清伟,王单单,王浩然,付秀虹..IGF-1基因克隆及其产物的测序鉴定[J].科技创新与应用,2024,14(16):7-11,5.基金项目
河南省自然科学基金(2223000420247) （2223000420247）
2023年河南省博士后科研资助项目(312145) （312145）
中央引导地方项目(Z20221343023) （Z20221343023）
河南省重点研发与推广专项(212102310813、222102310361) （212102310813、222102310361）
河南省医学科技攻关项目(LHGJ20221031) （LHGJ20221031）
漯河医学高等专科学校科技创新项目(2023ZD18) （2023ZD18）
