谢瓦氏曲霉间型变种flbA基因cDNA的全长克隆及序列分析

摘要

To explore the relationship between flb A gene structure and sexual reproduction, degenerate primers were designed from the conserved amino acids sequence of the FlbA protein, the full-length cDNA sequence of flbA gene was obtained from the Aspergillus chevalieri total RNA through RT-PCR (reverse transcription polymerase chain reaction ) and RACE (rapid amplification of cDNA ends) technology. The full sequence is 3060bp long, containing a 2295 bp open reading frame. Coding region of 764 amino acids with the molecular weight of 83181.19 and isoelectric point of 6.31 was an unstable hydrophilicity protein. The results of homologous analysis in GeneBank demonstrated that the sequence has the highest 77 % similarity with A. fumigatus on the nucleotide sequence. The amino acids encoding by the cDNA sequence of the flbA gene have a RGS ( Regulator of G protein Singaling ) and two DEP ( dishevelled, Egl-10, pleckstrin ) conserved domain. These domains were conserved in the other Aspergillus.%为进一步研究谢瓦氏曲霉间型变种中flbA基因的结构及其与有性发育的关系,根据FlbA氨基酸的保守序列设计简并引物,利用RT-PCR、RACE方法从谢瓦氏曲霉间型变种的总RNA中扩增出ftbA基因的cDNA序列.该序列全长3060 bp,包含2295bp的开放阅读框(open reading frame,ORF),预测编码一个由764个氨基酸组成的蛋白,该蛋白分子量为83 181.19,理论等电点为6.31,为不稳定的亲水蛋白.序列同源性为:该基因与烟曲霉(Aspergillus fumigatus)的发育调控子ftbA相似度,最高为77％,其编码的氨基酸含有一个RGS(Regulator ofG protein Singaling)和两个DEP(dishevelled,Egl-10,pleckstrin)保守域,与Aspergillus其他种的FlbA蛋白保守域一致.