克隆、原核表达和鉴定牛重组SHH蛋白  

Cloning, prokaryotic expression and identification of bovine recombinant SHH protein

作　　者：魏金销 谭旭信[2] 滑留帅[3] 安小鹏[4] 张晓建[5] 白跃宇 WEI Jinxiao;TAN Xuxin;HUA Liushuai;AN Xiaopeng;ZHANG Xiaojian;BAI Yueyu(Henan Province Breeding Cattle Genetic Properties Testing Center,Zhengzhou 450046,China;Henan Province Animal Products Quality Monitoring and Test Center,Zhengzhou 450008,China;Institute of Animal Husbandry and Veterinary Medicine,Henan Academy of AgriculturalSciences,Zhengzhou 450002,China;College of Animal Science and Technology,Northwest A&F University,Yangling 712100,China;Henan Institute of Science and Technology,Xinxiang 453003,China;Henan Institute of Animal Health Supervision,Zhengzhou 450008,China)

机构地区：[1]河南省种牛遗传性能测定中心,河南郑州450046 [2]河南省畜产品质量监测检验中心,河南郑州450008 [3]河南省农业科学院畜牧兽医研究所,河南郑州450002 [4]西北农林科技大学动物科技学院,陕西杨凌712100 [5]河南科技学院,河南新乡453003 [6]河南省动物卫生监督所,河南郑州450008

出　　处：《畜牧与兽医》2019年第5期6-10,共5页Animal Husbandry & Veterinary Medicine

基　　金：国家高层次人才特殊支持计划"万人计划"(2069999)

摘　　要：Sonic Hedgehog(SHH)是Hedgehog家族的一员,其不仅参与胚胎发育的基本过程,同时也参与脂肪生成和肌肉生成。SHH在动物肌肉和脂肪沉积上的双向作用提示它是调控肌内脂肪含量(IMF)的重要候选基因。本研究中,使用重叠PCR法克隆了去除信号肽序列的牛SHH(btSHH)基因,然后在大肠杆菌BL21中诱导表达了btSHH蛋白。使用镍亲和层析试剂盒纯化了原核表达的btSHH,并用Western blot验证了目标蛋白的正确性。将纯化后的btSHH添加到前脂肪细胞3T3-L1的培养基中,并诱导细胞分化后表明,重组btSHH蛋白能够显著地抑制脂肪沉积过程。本研究结果为未来肉牛生产中应用btSHH提供理论基础。Sonic Hedgehog(SHH),a member of the Hedgehog family,not only participates in the fundamental processes of embryonic development,but is also involved in adipogenesis and myogenesis.The dual effects SHH on the development of adipose and muscle tissues indicate that it might be a candidate regulatory factor for intramuscular fat(IMF) deposition in farm animals.In the present study,the Bos taurus SHH(btSHH) gene without the signal peptide sequence was cloned by overlap PCR,and then the btSHH protein was induced to express on E.coli BL21 and was purified by Ni-affinity chromatography.The purified btSHH was verified by western blot and then added to the culture medium of pre-adipocyte 3T3-L1 during the induced differentiation process.The Oil Red O staining showed that the recombinant btSHH could significantly inhibit the adipogenesis.This research might provide a basis for the application of btSHH in beef cattle productions in the future.

关 键 词：SHH蛋白 克隆 重叠PCR 原核表达 功能鉴定 

分 类 号：S813[农业科学—畜牧学]