[Cloning and characterization of glyceraldehyde-3-phosphate dehydrogenase gene from Chaetomium globosum].
The amino acid sequence of Glyceraldehyde 3-phosphate dehydrogenase (GAPDH) gene from Neurospora crassa (XP-327967) and Colletotrichum lindemuthianu (P35143) were subjected to local tBlastn searching against the ESTs local datebase of Chaetomium globosum. The full length cDNA sequence of 1240bp encoding GAPDH gene with an open reading frame of 1014bp and encoding 337 amino acids was obtained. The protein molecular weight was 36.1kD. The DNA sequence of GAPDH gene was obtained through PCR amplification with specific primers of cDNA 5' and 3' untranslated region. The analysis of DNA sequence indicated that GAPDH gene have 3 exons and 2 introns. The BlastP analysis revealed that amino acids sequence of GAPDH gene from C. globosum shared 95% high similarity with Podospora anserin and 87% low similarty with Aspergillus oryzae. A transgenic yeast harboring C. globosum GAPDH was generated under the control of a constitutively expressed GAL promoter. The results from biofunctional analyses of GAPDH yeast transformants showed that GAPDH yeast transformants had significantly higher resistance to Na2 CO3 and heat stresses. The cDNA, DNA and deduced amino acid sequence of GAPDH gene were accepted by GenBank (accession numbers: AY522719, AY593253, AAS01412).
Liu
Zhi-Hua
ZH
Department of Life Science and Engineering, Harbin Institute of Technology, Harbin 150001, China. liuzhihua1029@yahoo.com.cn
Yang
Qian
Q
D001483:Base Sequence; D002254:Carbonates / Q000494:pharmacology; D002597:Chaetomium / Q000201:enzymology* / Q000235:genetics; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000302:isolation & purification; D005987:Glyceraldehyde-3-Phosphate Dehydrogenases / Q000737:chemistry / Q000235:genetics*; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction; D017422:Sequence Analysis, DNA; D014170:Transformation, Genetic; D015003:Yeasts / Q000235:genetics
D002254:Carbonates; D018076:DNA, Complementary; C005686:sodium carbonate; D005987:Glyceraldehyde-3-Phosphate Dehydrogenases