鸭CYP7_α1基因克隆、序列分析及组织表达研究

cDNA Cloning,Sequence Analysis and Tissue Expression of Duck CYP7_α1 Gene

旨在克隆鸭胆固醇7α羟化酶1(CYP7α1)基因,并进一步探讨该基因的结构与功能,揭示其组织特异性表达规律。本研究以樱桃谷鸭为材料,运用同源序列克隆结合RACE技术,对鸭CYP7α1基因的cDNA全长进行克隆,并对其进行生物信息学分析及组织表达的研究。结果表明:鸭CYP7α1基因cDNA全长2 351bp,最大开放阅读框(ORF)1 539bp(114～1 652bp),共编码512个氨基酸;鸭CYP7α1氨基酸与鹅的同源性最高(97.5%),其次是鸡(92.8%)、人(67.3%)、猪(66.5%)、兔(65.7%)、大鼠(65.5%)、小鼠(65.5%)和牛(65.1%);经预测鸭CYP7α1蛋白可能含有1个P450超家族结构域;鸭CYP7α1基因在肝脏中呈高丰度表达,在其他9个组织中均未检测到或检测到少量表达,表明该基因的表达具有组织特异性。

This study was carried out to obtain and analyze sequence of duck CYP7αl gene, and to reveal the expression of CYP7αl mRNA in different tissues of duck. Based on other species mR- NA sequences, the cDNA of CYP7αl gene in duck was amplified successfully by using RACE- PCR. The structure and the function of the duck CYP7αl gene were analyzed by bioinformatics methods. The mRNA abundance of duck CYP7αl gene was tested in 10 tissues. The result showed that the cDNA of duck CYP7α1 gene was 2 351 nucleotides in length with an open reading frame （ORF） 1 539 bp（114-1 652 bp） in length and encoded 512 amino acids. The amino acid ho- mology of duck CYP7αl were 97.5%, 92.8%, 67.3%, 66.5%, 65.7%, 65.5%, 65.5% and 65.1% comparing to that of the goose, chicken, human, pig, rabbit, rat, mouse and cattle, re- spectively. The CYP7αl amino acid sequence of duck contained a P450 domain. The mRNA ex- pression level of duck CYP7al gene was high in liver and was not detected in other 9 tissues, the mRNA expression of duck CYP7al gene was tissue specific.