%0 Journal Article
%T 一株尖孢镰刀菌细胞色素P450氧化酶基因的克隆及生物信息学分析<br>Molecular cloning and bioinformatics analysis of cytochrome P450 oxidase gene from fusariumoxysporum
%A 单丽红
%A 姜冬冬
%A 赵沙沙
%A 黄佳佳
%A 朱
%A 丽
%A 刘宏民
%J 郑州大学学报（医学版）
%D 2018
%R 10.13705/j.issn.1671-6825.2017.05.123
%X 目的:克隆一株能高效转化合成屈螺酮的重要中间体(7α,15α-二羟基DHEA)的尖孢镰刀菌参与反应的关键酶细胞色素P450氧化酶(CYP)基因CYP6003A1。方法:根据尖孢镰刀菌转录组得到CYP基因,自主设计引物,运用Trizol法提取DHEA诱导48 h的尖孢镰刀菌的总RNA,通过PCR扩增尖孢镰刀菌CYP基因并进行基因测序; 运用生物信息学分析得到的cDNA序列。结果:成功扩增出全长3 459 bp的cDNA片段,将CYP6003A1基因的氨基酸序列通过NCBI上的BLAST进行在线比对,对照结果显示与CYP6003A1相似度前10个基因有3个是CYP基因,其余均是与CYP基因相关的基因。CYP6003A1基因生物信息学分析结果显示,该基因片段编码691个氨基酸的多肽。结论:从尖孢镰刀菌中成功克隆了CYP6003A1基因并进行了生物信息学分析。<br>Aim: To clone the key enzyme gene, CYP6003A1, which was involved in the reaction of fusariumoxysporum, which could transform drospirenone into an important intermediate(7α, 15α-diOH-DHEA).Methods: Primers were designed according to the CYP gene of fusariumoxysporum. The total RNA of fusariumoxysporum was extracted by the method of Trizol extraction from fusariumoxysporum induced by DHEA for 48 h, and the CYP gene of fusariumoxysporum was amplified by PCR and sequenced. The bioinformatics were analyzed.Results: A full-length cDNA fragment of 3 459 bp was successfully amplified. After sequencing, the amino acid sequence of the CYP6003A1 gene was screened by BLAST on NCBI. The results showed that the first 10 genes were similar to CYP6003A1, among them, three were CYP gene, and the others were genes related to CYP gene. The result of bioinformatics analysis showed that the gene fragment encoded 691 amino acids polypeptide.Conclusion: CYP6003A1 gene has been successfully cloned from fusariumoxysporum and analyzed by bioinformatics
%K 7α
%K 15α-二羟基DHEA
%K 细胞色素P450氧化酶
%K 尖孢镰刀菌
%K 生物信息学&lt
%K br&gt
%K 7α
%K 15α-diOH-DHEA
%K cytochrome P450 oxidase
%K fusariumoxysporum
%K bioinformatics
%U http://jms.zzu.edu.cn/oa/darticle.aspx?type=view&id=201801009