擎天凤梨GAPDH基因的克隆及序列分析

管家基因GAPDH(glyceraldehyde-3-phosphate dehydrogenase)常被用作定量、半定量PCR试验的内参,以确定目标基因的相对表达量。基于前期从全长cDNA文库中获得GAPDH基因的EST单克隆,进行Primer Walking测序,获得一个GAPDH的全长cDNA序列,序列长1 790 bp,编码421个蛋白氨基酸,命名为GoGAPDH1。采用Blast,ExPASy,ProtParam,SPOMA,Find Conserved Domains(NCBI),ClustalX, MEGA等生物信息学软件对cDNA 序列、氨基酸序列、保守区、亲缘关系等特征进行分析。结果表明，GoGAPDH1与玉米的细胞质GAPDH基因源性最高，为82%。初步推断GoGAPDH1可能是一个胞质型GAPDH。

Cloning and sequence analysis of GAPDH gene from Guzmania

House keeping gene GAPDH(glyceraldehyde-3-phosphate dehydrogenase) was often used as inner control in quantitative or semi-quantitative PCR test to determine relative expressive amount of target genes. Based on EST monoclones of GAPDH gene obtained from Guzmania full length cDNA library, its full length cDNA sequence was obtained by Primer Walking sequencing. The gene, designated GoGAPDH1, was 1 790 bp in length encoding 421 amino acids. Some characters involving in cDNA sequence, amino acid sequence, conservative domain and genetic relationship were analyzed by Blast, ExPASy, ProtParam, SPOMA, Find Conserved Domains(NCBI), ClustalX and MEGA softwares． The results showed that GoGAPDH1 shared 82% similarity with cytoplasmic GAPDH in maize, which indicated that GoGAPDH1 might be a cytoplasmic GAPDH.