摘要
GM-CSF在机体免疫系统中发挥重要的免疫调节作用。用PHA与LPS体外联合刺激猪外周血单个核细胞,采用RT-PCR技术成功克隆了猪GM-CSF基因。序列分析表明,该序列与NCBI/GenBank上登载的序列有两个碱基不同;将猪与人、牛、羊、狗、猫、鼠等动物的GM-CSF基因序列相比较,其一致性分别为82.1%8、5%、88.1%、82.8%、83.7%、70.5%,在氨基酸水平上一致性为70.3%7、0.9%7、8.1%、70.3%7、1.9%、54.4%。然后利用生物信息学和分子生物学软件,对猪GM-CSF的结构进行预测,结果表明,猪GM-CSF为一结构松散的球状蛋白分子。猪GM-CSF基因的成功克隆及其编码蛋白结构的预测为开发高效、低毒且性质稳定的猪GM-CSF产品奠定了基础。
GM-CSF plays very important role in immune system regulation. We isolated porcine peripheral lymphocytes cells and costimulation it with PHA and LPS.We then extracted total RNA from the cells and amplified GM-CSF gene using RT-PCR.The obtained fragment was ligated into pGEM-T easy vector and sequenced. The result showed that the sequence had 2 nucleotides difference with the sequence of GM-CSF gene published in GenBank. sequence analysis shows 82 % identity with human, 85 % with bovine, 88 % with ovine,83 % with canine,84 % with feline,and 71% with murine GM-CSF, and 70.3 % ,70.9 % ,78.1% ,70.3 % ,71.9 % and 54.4 % at the GM-CSF mature protein amino acid level. The protein structure prediction showed that the GM-CSF protein is a loose globe protein. These results pave the way for future developments on the GM-CSF product.
出处
《中国预防兽医学报》
CAS
CSCD
北大核心
2005年第6期474-478,482,共6页
Chinese Journal of Preventive Veterinary Medicine
基金
国家高技术发展计划("863")资助项目
项目编号:2003AA24111
甘肃省自科学基金资助项目
项目编号:ZX031-A25-045-D
