黑灵芝免疫调节蛋白基因的克隆和生物信息学分析

黑灵芝(Ganoderma astum)是灵芝家族中一个重要成员,克隆和分析黑灵芝免疫调节蛋白基因,可为进一步了解真菌免疫调节蛋白在种群中的分布打下基础。以黑灵芝为材料,采用同源克隆方法,对真菌免疫调节蛋白(FIPs)基因进行PCR扩增,并对其核苷酸编码序列进行了生物信息学分析。结果表明:黑灵芝真菌免疫调节蛋白基因序列包含336bp,可编码111个氨基酸残基,命名为FIP-gas。FIP-gas分子量为12.4kD,理论等电点为4.80,符合FIPs的特性,是FIPs家族一新成员。对FIP-gas进行序列分析发现其氨基酸序列具有FIPs的保守序列模式,与紫灵芝(G.japoncium)、紫芝(G.sinensis)、小孢灵芝(G.microsporum)、赤灵芝(G.lucidum)、松杉灵芝(G.tsugae)、金针菇(Flammulina velutipes)及草菇(Volvariella volvacea)的FIP序列的同源性分别为99%、98%、87%、86%、86%、61%、55%。系统进化树分析表明黑灵芝的FIP与紫芝的FIP亲缘性最高,与金针菇和草菇的FIP亲缘性相对较远。该研究为进一步了解免疫调节蛋白在灵芝种群中的分布提供了较详细的数据。

Cloning and Bioinformatics Analysis of Fugal Immunomodulatory Protein Gene from Ganoderma astum

A fugal immunomodulatory protein(FIP) gene sequence was amplified from the genomic DNA of Ganoderma astum by PCR with a pair of specific primers,designed as FIP-gas.The sequence analysis showed that the fragment contains a full encoding region of 336 bp encoding 111 amino acid residues with a molecular mass of 12.4 kD.This deduced protein has a pI of 4.80.The result of sequence analysis indicated that FIP-gas was a new member of FIP family.The deduced amino acid sequence of FIP-gas shares high similarity with other members of FIP family,such as FIP-gja(G.japoncium),FIP-gsi(G.sinensis),FIP-gmi(G.microsporum),FIP-glu(G.lucidum),FIP-gts(G.tsugae),FIP-fve(Flammulina velutipes) and FIP-vvo(Volvariella volvacea),which were found to be 99%,98%,87%,86%,86%,61% and 55% identical,respectively.Meanwhile the phylogenetic relationships were analyzed using the neighbor joining method;The phylogenetic tree shows that FIP-gas has a close relationship with FIP-gsi and FIP-gja.This study provides an important foundation for further study of FIPs′ community distribution.