3'RACE法扩增谷氨酰胺合成酶基因及其生物信息学分析

摘要

以抽提得到的黑曲霉mRNA为模板,根据GenBank上检索的谷氨酰胺合成酶基因mRNA序列,设计特异性引物,进行3 ' RACE PCR扩增,并在NCBI网站进行生物信息学分析.结果扩增得到长约1 kb的DNA片段,测序结果表明,所获得的DNA序列与gene bank上检索到黑曲霉产谷氨酰胺合成酶基因的同源性大于97％,与其他曲霉产谷氨酰胺合成酶的基因同源性大于50％.经开放阅读框软件分析发现其具有2个外显子,其中1个外显子的氨基酸序列与黑曲霉产GS同源性为100％.%On the basis of GS mRNA and specific primer, Using 3'RACE PCR amplified the gene sequence and analyzed some bioinformatics information of the sequence in NCBI web. GS DNA fragment was about 1. 0 kb. The sequence blast showed that the homology of fragment with GS gene of Asp. niger in GenbBank was more than 97 % and with other Asp were more than 50%. Open reading frame analyzed that it included 2 exon, one of which had 100% homology with GS of Asp. niger.