Biotechnology Bulletin ›› 2016, Vol. 32 ›› Issue (7): 131-137.doi: 10.13560/j.cnki.biotech.bull.1985.2016.07.020

• Orignal Article • Previous Articles     Next Articles

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

ZHANG Hong ZHENG Rong   

  1. College of Life Sciences and Technology,Inner Mongolia Normal University,Hohhot 010022
  • Received:2015-12-25 Online:2016-07-25 Published:2016-07-25

Abstract: Designing the primers based on the conserved sequences of several fungal actin genes,the full-length cDNA sequence of γ-actin gene from Leucocortinarius bulbiger(Lb-act)was cloned using RT-PCR and RACE. The full-length of Lb-act cDNA 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 high similarity 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 research confirmed the reliability that actin gene could be used as intermolecular standard.

Key words: Leucocortinarius bulbiger, actin gene, cDNA, clone