摘要
选用猪为研究对象,以可卡因苯异丙胺调控转录肽(CART)作为影响猪繁殖机能的多肽,依据NCBI上登录的CARTmRNA序列设计3对特异性引物,用RT-PCR方法从下丘脑内扩增出CART mRNA的部分序列并进行序列分析;再根据扩增出的序列设计一对带酶切位点的特异性引物,用PCR方法扩增出含EcoRⅠ/HindⅢ酶切位点的CART mRNA的全编码序列(CDS)区序列.结果表明:猪下丘脑CART mRNA的CDS区全长367 bp,CART基因在下丘脑上有表达,并成功构建pEG-32a-CART原核表达载体.
Designing three pairs specific primers based on the CART mRNA sequence included in NCBI GenBank,CART mRNA sequences were amplified via RT-PCR from porcine hypothalamus tissue,and were analyzed.According to the amplified sequence,we designed a pair of expressing primers,amplified the whole coding sequence（CDS） domain of CART including EcoRⅠ/HindⅢ digestion sites.The results showed that： the length of CDS in porcine hypothalamus CART mRNA is 367 bp,CART gene can express in the hypothalamus,and it had successfully constructed prokaryotic expression vector of pEG-32a-CART.
出处
《福建农林大学学报（自然科学版）》
CSCD
北大核心
2011年第6期610-613,共4页
Journal of Fujian Agriculture and Forestry University:Natural Science Edition
基金
农业部"948"项目(2010-Z43)
山西省留学基金
山西省引进人才专项基金项目(2008080)
山西省国际科技合作项目(2010081002)
