版纳微型猪近交系细胞色素P450家族基因CYP1A1克隆及生物信息学分析

Cloning and Bioinformatics Analysis of CYP1A1 of Cytochrome P450 Family from Banna Mini-pig Inbred Line

【目的】克隆版纳微型猪近交系(Banna mini-pig inbred line,BMI)细胞色素P450家族基因CYP1A1的编码区序列,并对其进行生物信息学分析。【方法】利用RT-PCR方法获得CYP1A1基因的全长CDS序列,运用生物信息学对其核酸及蛋白序列进行分析,构建系统进化树。【结果】获得的BMI CYP1A1 CDS长度为1551 bp,编码516个氨基酸,蛋白质分子量为58.39 ku,等电点为7.83,位于内质网的概率是94.1%。CYP1A1蛋白质含有1个细胞色素P450半胱氨酸血红素配体位点,1个N-豆蔻酰化位点,1个酰胺化作用位点,2个N-糖基化位点、6个蛋白酶C磷酸化位点和5个蛋白激酶Ⅱ磷酸化位点。11个物种构建的系统进化分析表明,BMI(猪)CYP1A1与牛和山羊的关系最近。【结论】以上结果将为进一步研究版纳微型猪近交系CYP1A1基因功能奠定基础。

[ Purpose] Our purpose was to clone sequence and analyze the character of the cytochrome P450 family 1 subfamily A member 1 gene （CYP1A1） from Banna mini-pig inbred line （BMI）. [ Methods] The complete CDS of CYP1A1 gene was cloned using RT-PCR method from BMI. Nucleo- tide and protein sequences of CYPIAI were utilized to cany out bioinformatics analysis and constructthe phylogenetic tree. [ Results] The CDS length of BMI CYPIA1 is 1 551 bp in length which encodes a protein of 516 amino acids with molecular weight （Mw） 58.39 ku, and isoelectric point （pI） 7.83, and locates in the endoplasmic reticulum with 94. 1% reliability. CYP1AI protein contains a cyto- chrome P450 cysteine heme-iron ligand signature, an N-myristoylation site, a aidation site, two N-gly- cosylation sites, six protease C phosphorylation sites and five protein kinase II phosphorylation sites. Phylogenetic analysis of 11 species showed that CYP1 A1 of BMI （pig） had the closest relationship with these of cattle and goat. [ Conclusion] The results will lay a foundation for further study on CYP1A1 gene function in BMI.