摘要
目的探讨克隆高原藏羚羊胱硫醚-γ-裂解酶(CSE)基因编码区并检测其在成年高原藏羚羊组织中的表达,同时探讨高原藏羚羊低氧适应的分子生物学机制。方法从高原藏羚羊组织中提取总RNA,通过逆转录聚合酶链反应(RT-PCR)获得高原藏羚羊cDNA,并进行鉴定和测序。结果将含有目的片段克隆后经测序和Blast分析,结果显示其部分编码序列与GenBank中牛CSE蛋白基因序列同源性96.47%,表明本实验所克隆的序列为CSE蛋白基因。根据已知人、褐家鼠、小鼠、野猪、食蟹猴、家牛CSEcDNA序列和Pnaman软件设计高原藏羚羊cse基因的引物。结论 CSE mRNA可能在高原藏羚羊机体较为广泛的区域中发挥着作用,同时为高原低氧适应相关基因的研究提供了实验依据。
Objective To identify the Cystathionine-γ-Lyase（CSE） genes coding sequences molecular cloning, exam the tissues expression spectrums and discuss the hypoxic adaptations mechanisms in Tibetan antelope. Methods The total RNA was extracted, and the eDNA was captured by reverse transcription RT-PCR, then identified, se- quenced and cloned. Results There was 96.47%homology between the Tibetan antelope gene fragment containing the purpose gene and the cattle gene in gen banks,thus the result mean the gene which cloned before was CSE gene protein. The length of the CSE gene protein was been detected by designing primers according to the human,mous, wild boar, cattle CSEcDNA sequences,and the CSE gene primers of tibetan antelope which tesied by Pnaman. Conclu- sion CSE gene protein might play an important role in the body of the Tibetan anteplope,which provide experiment basis to the gene study about adaptation in high altitude hypoxia environment, in the future.
出处
《检验医学与临床》
CAS
2013年第23期3099-3101,共3页
Laboratory Medicine and Clinic
基金
国家"973"资助项目(2006CB504100)
