摘要
CCDC174被发现是鸡原始生殖细胞(Primordial germ cells,PGCs)关键调控基因C2EIP的互作基因,在PGCs起源、发育和分化中发挥重要作用。试验克隆CCDC174基因全长并对其进行生物信息学分析,构建过表达和干扰载体进行检测。结果表明,CCDC174主要定位于细胞核中,有DUF4078、PTZ00121和PABP 3个功能域,存在磷酸化修饰位点且该基因在鸟类中保守性较高;酶切结果表明CCDC174载体构建成功,q RT-PCR和Western blot结果进一步表明构建的载体可有效干扰和过表达CCDC174,为后期对CCDC174功能验证和机制探究提供参考依据和技术手段。
CCDC174 was found to be the interaction gene of the key regulatory gene C2EIP in primordial germ cells(PGCs) of chicken, which played an important role in the origin, development and differentiation of PGCs. In this study, the full length of CCDC174 gene was cloned and analyzed by bioinformatics, and its overexpression vector and interference vector were constructed and detected.The results showed that CCDC174 was mainly located in the nucleus and had three functional domains of DUF4078, PTZ00121 and PABP. There were phosphorylation modification sites and CCDC174 was highly conserved in birds. Enzyme digestion results showed that CCDC174 vector was successfully constructed, and qRT-PCR and Western blot results further indicated that the constructed vector could effectively interfere with and overexpress CCDC174, which provided a reference basis and technical means for functional verification and mechanism exploration of CCDC174 at the later stage.
作者
周静
夏钱
孙国兴
靳锴
牛英杰
孙红艳
左其生
张亚妮
李碧春
ZHOU Jing;XIA Qian;SUN Guoxing;JIN Kai;NIU Yingjie;SUN Hongyan;ZUO Qisheng;ZHANG Yani;LI Bichun(School of Animal Science and Technology,Yangzhou University,Yangzhou Jiangsu 225009,China;Joint International Research Laboratory of Agriculture and Agri-Product Safety of Ministry of Education,Yangzhou Jiangsu 225009,China)
出处
《东北农业大学学报》
CAS
CSCD
北大核心
2022年第5期42-50,79,共10页
Journal of Northeast Agricultural University
基金
国家自然科学基金项目(32002158)
中国博士后科学基金项目(2020M681746)。
