偏肿革裥菌C2H2型转录因子SFP1基因克隆、生物信息学及表达分析

Cloning,Bioinformatics and Expression Analysis of SFP1 Gene of C2H2 Zinc Finger Transcription Factor in Lenzites gibbosa

以L.gibbosa菌丝体总RNA反转录的cDNA为模板,通过RT-PCR克隆得到一个C2H2型锌指转录因子的cDNA序列。对该基因进行基因克隆、生物信息学分析,并利用实时荧光定量法对木屑处理下该基因的表达进行分析。通过保守结构域预测与结构分析发现该基因为裂指蛋白1类(SFP1)C2H2型锌指转录因子,具有2个C2H2家族的保守结构域,将该基因命名为Lg-sfp1。该基因CDS全长1947 bp,编码一个由648个氨基酸组成的亲水性膜内蛋白,蛋白大小为68.27 ku。通过SWISS-MODEL预测的三级空间结构发现两个完整的C2H2结构。木屑处理下Lg-sfp1基因表达比无木屑处理下表达量低,推断木屑促进了L.gibbosa氧化应激反应,当氧化应激反应大量发生时,参与氧化应激反应负调控的sfp1基因表达受到抑制,为后续研究偏肿革裥菌sfp1基因在木材降解,特别是氧化应激反应中的功能提供了理论基础。

A C2H2 zinc finger transcription factor was cloned by RT-PCR using cDNA reverse transcription of total RNA from L.gibbosa strain.The gene was cloned andanalyzedin bioinformatics,and the expression of the gene under sawdust treatment was analyzed by Quantitative Real-time PCR method.Through conservative domain prediction and structural analysis,the gene is a Split Zinc-finger protein 1(SFP1)C2H2 transcription factor with two C2H2 family conserved domains.The gene was named Lg-sfp1.The CDS length of the gene is 1947 bp,encoding a hydrophilic membrane protein composed of 648 amino acids,with a protein size of 68.27 ku.Through thethree-dimensional structure predicted by SWISS-MODEL,two complete C2H2 structures were found.The expression level of Lg-sfp1 gene was lower under sawdust treatment than that without sawdust treatment,inferring that sawdust promoted oxidative stress response of Lg-sfp1 gene expression involved in negative regulation of oxidative stress response was inhibited when a large number of oxidative stress responses occurred.