单酶切cDNA-AFLP改良法分离草菇冷诱导基因

ISOLATION OF COLD INDUCED GENES BY MODIFIED cDNA-AFLP WITH SINGLE ENZYME DIGESTION FROM VOLVARIELLA VOLVACEA

采用单酶切cDNA-AFLP改良法克隆分离了十条草菇冷诱导基因片段VC7、VC8、VC9、VC10、VC11、VC12、VC13、VC14、VC15、VC16。DNA序列测定结果表明,克隆分离的十条DNA片段分别为504bp、549bp、555bp、301bp、189bp、602bp、605bp、834bp、703bp、1085bp。同源性BLAST程序搜索结果显示,除VC14与粗糙脉孢菌Neurosporacrassa的钙磷型ATP酶基因有较高的同源性外,其余九条DNA片段目前都没有搜索到同源的核酸序列;而其翻译的蛋白质序列,除了VC11和VC15目前未能搜索到同源的蛋白质序列外,其余八条DNA片段翻译的蛋白质序列与粗糙脉孢菌Neurosporacrassa、裂殖酵母Schizosaccharomycespombe、链霉菌Streptomycesavermitilis等的有关蛋白质序列具有较高的同源性。

Ten cold induced gene fragments VC7, VC8, VC9, VC10, VC11, VC12, VC13, VC14, VC15, VC16 were isolated from Volvariella volvacea by modified cDNA-AFLP with single enzyme digestion. DNA sequence results revealed that these DNA fragments had 504bp, 549bp, 555bp, 301bp, 189bp, 602bp, 605bp, 834bp, 703bp, 1085bp respectively. BLAST search results showed that VC7, VC8, VC9, VC10, VC11, VC12, VC13, VC15, VC16 had no significant homologous nucleotide sequences found in the databases yet except that VC14 was highly homologous with Neurospora crassa partial nca-1 gene for calcium P-type ATPase. Translated protein sequences of VC7, VC8, VC9, VC10, VC12, VC13, VC14, VC16 had high similarities to some corresponding protein sequences of Neurospora crassa, Schizosaccharomyces pombe and Streptomyces avermitilis, while translated protein sequences of VC11 and VC15 had no significant homologous protein sequences found in the databases yet.