摘要
核基质结合蛋白SATB1调控染色质的空间结构,参与真核生物基因表达。本论文通过PCR获得SATB1目的基因,然后与载体mCherry-C1同时用EcoRⅠ和KpnⅠ双酶切后,用T4连接酶进行连接。连接质粒转入感受态宿主细胞并在Kana抗性的LB培养基中培养。分离获得Kana抗性单克隆菌株并进行扩大培养后提取重组质粒。对重组质粒进行双酶切验证和DNA测序检测。本论文成功构建mCherry-SATB1真核表达质粒,为后续研究SATB1的功能机制奠定基础。
Nuclear matrix binding protein SATB1 participates in eukaryotic gene expression by regulating the spatial structure of chromatin.In this study,the SATB1 gene fragment was amplified by PCR.The PCR product and mCherry-C1vector were digested with EcoRⅠand KpnⅠ,and then connected with T4 ligase.The constructed plasmid was transferred into competent host cells DH5αand cultured on Kana+LB medium,and the single colony was selected and cultured.The constructed plasmid was verified by double enzyme digestion and DNA sequencing.It is proved that the eukaryotic expression plasmid mCherry-SATB1 was successfully constructed and can be used for further research of SATB1 function.
作者
齐文靖
杨壮
韩国军
QI Wen-jing;YANG Zhuang;HAN Guo-jun(College of Life Sciences,Changchun Normal University,Changchun 130032,China)
出处
《长春师范大学学报》
2023年第4期111-114,共4页
Journal of Changchun Normal University
基金
吉林省教育厅项目“肌动蛋白协同SATB调控炎症反应中相关基因表达的机制研究”(JJKH20200825KJ)
吉林省科技厅项目“BRCA2在肿瘤迁移中的作用机制研究”(YDZJ202201ZYTS661)
长春师范大学项目“先天免疫反应中肌动蛋白调控SATB1的作用机制研究”(长师大自科合字[2019]第017号)。
