球根白丝膜菌γ-actin基因的克隆及表达分析  

Cloning and Expression Analysis of γ-actin Gene from Leucocortinarius bulbiger

作　　者：张虹[1] 峥嵘[1] 

机构地区：[1]内蒙古师范大学生命科学与技术学院,呼和浩特010022

出　　处：《生物技术通报》2016年第7期131-137,共7页Biotechnology Bulletin

基　　金：国家自然科学基金项目(31060110;31360125);内蒙古自然科学基金项目(2012MS0526)

摘　　要：根据真菌肌动蛋白(actin)基因保守区序列设计引物,用简并PCR法和RACE技术分离得到球根白丝膜菌(Leucocortinarius bulbiger)γ-肌动蛋白基因(Lb-act)的全长cDNA序列。该序列全长为1 357 bp,包含一个1 137 bp的开放阅读框(ORF),编码378个氨基酸,5'端非翻译区(5'UTR)92 bp,3'UTR长度128 bp。Port Param软件在线分析结果表明,该cDNA所编码的蛋白质理论等电点为5.12,相对分子质量为95.022 kD,具有真菌γ-actin基因3个保守特征序列。Blast同源性检索结果表明,Lb-act氨基酸序列与担子菌肌动蛋白序列有较高的相似性,其与双色蜡蘑的肌动蛋白氨基酸序列的亲缘关系最近。Lb-act基因在不同碳源及磷水平培养条件下表达量基本一致,验证了该基因作为分子内标的可靠性。Designing the primers based on the conserved sequences of several fungal actin genes,the full-length c DNA sequence of γ-actin gene from Leucocortinarius bulbiger（Lb-act）was cloned using RT-PCR and RACE.The full-length of Lb-act c DNA sequence was 1 357 bp,consisting of a 1 137 bp open reading frame（ORF）,encoding a protein of 378 amino acids,a 5＇-UTR with 92 bp and a 3＇-UTR with 128 bp.The online analysis by Port Param software revealed that the putative amino acids had an isoelectric point of 5.12,a molecular weight of 95.022 kD,and 3 highly conserved regions of fungal γ-actin gene.Blast homology search indicated that the sequences of Lb-act amino acid had a highsimilarity with the sequences of Basidiomycetes actin,and it had the closest relationship with the amino acid sequence of Laccaria bicolor actin.In culture conditions of different carbon and phosphorus levels,the expression levels of Lb-act were almost the same,thus this researchconfirmed the reliability that actin gene could be used as intermolecular standard.

关 键 词：球根白丝膜菌 肌动蛋白 CDNA 克隆 

分 类 号：Q93[生物学—微生物学] Q78