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首页> 外文期刊>African Journal of Biotechnology >Molecular cloning of endochitinase 33 (ECH33) gene from Trichoderma harzanium
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Molecular cloning of endochitinase 33 (ECH33) gene from Trichoderma harzanium

机译：哈茨木霉内切酶33（ECH33）基因的分子克隆

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This?study was conducted to screen for the presence of?ech33 gene in 80 isolates of?Trichoderma.?Furthermore, using gene specific primers,?ech33 gene were cloned into pTZ57R/T from?T. harzanium?IABT1068. The clone was confirmed through PCR amplification and restriction analysis. The clones were sequenced and analyzed for homology at nucleotide and protein level to find out conserved domain of protein. Gene encoding endochitinase from both species have 96 and 95% homology with reported sequence both at nucleotide and protein level.?The cloned?ech33 has a size of 1159 bp, of which 9 bp corresponds to the 5' untranslated region, with a?650 bp?open reading frame.?The amino acid sequence of gene has?signal peptide sequence ranges from 1 to 19.?The nucleotide sequence analysis using GENETOOL software revealed presence of?three exon and four introns, and?has unique restriction sites for?HindIII,?BamHI?and?SalI,?at 881, 308 and 485 positions, respectively.
机译：本研究旨在筛选80种木霉属细菌中ech33基因的存在。进而，使用基因特异性引物将ech33基因从ΔT克隆到pTZ57R / T中。 za？IABT1068。通过PCR扩增和限制性酶切分析确认克隆。对克隆进行测序，并在核苷酸和蛋白质水平上分析同源性，以找出蛋白质的保守结构域。来自这两个物种的编码内切壳多糖酶的基因在核苷酸和蛋白质水平上与报道的序列具有96％和95％的同源性。克隆的echech33的大小为1159bp，其中9bp对应于5'非翻译区，其中a？650 bp的开放阅读框。该基因的氨基酸序列的信号肽序列范围为1至19。使用GENETOOL软件进行的核苷酸序列分析显示，存在3个外显子和4个内含子，并且具有唯一的限制性酶切位点。 HindIII，BamHI和SalI分别位于881、308和485位。

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