Cloning and Characterization of Glyceraldehyde-3-Phosphate Dehydrogenase From Orchid Cymbidium goeringii)
The total RNA of Cymbidium goeringii (C. goeringii) leaves was extracted with Trizol. With primers designed based on the sequence of gtyceraldehyde-3phosphate dehydrogenase (GAPDH) from NCBI, a fulllength cDNA of GAPDH was cloned from C goeringii by RT-PCR combined with 3RACE and 5RACE. The newly cloned GAPDH gene was designated as CgGAPDHl (JQ978220). Sequence analysis shows that CgGAPDHl is 1 424 bp in length, contains a single open reading frame from 168 bp to 1 190 bp, encodes proteins with 340 amino acids. CgGAPDHl showed 99％ similarity with GAPDHS (JN177726.1). Phylogenetic analysis based on the sequence of GAPDH showed that C. goeringii had closely relation with Triticum aestivum and Lilium longiflor. Bioinformatics analysis indicated that CgGAPDHl protein is soluble without transmembrane domain. Domain analysis revealed the proteins contain two conserved domains, named as NADBRossmann and Gp_dh_C. Homology modeling of the GAPDH firmed that the polypeptides was highly conserved. RT-PCR indicated CgGAPDHl can be used as internal control gene.
Glyceraldehyde-3-phosphate dehydrogenase cloning bioinformatics RACE
Yunfang Tian Xiuyun Yuan Ning Wang Jinle Su Bo Cui
Institute of Biotechnology,Zhengzhou Normal University Zhengzhou, China College of Forestry,Henan Agricultural University Zhengzhou, China
国际会议
哈尔滨
英文
423-425
2012-06-16（万方平台首次上网日期，不代表论文的发表时间）