首页> 外国专利> Heterologous expression of fungal polyketide synthetic gene in yeast and a method of preparing a compound produced by a protean encoded by the polyketide synthetic gene by the heterologous expression

Heterologous expression of fungal polyketide synthetic gene in yeast and a method of preparing a compound produced by a protean encoded by the polyketide synthetic gene by the heterologous expression

机译:真菌聚酮化合物合成基因在酵母中的异源表达以及通过异源表达制备由聚酮化合物合成基因编码的蛋白的蛋白产生的化合物的方法

摘要

The present invention relates to a method of removing an intron contained in a gene from a eukaryotic gene, and linking only the exon sequences to prepare an expression vector comprising the linked sequences. Specifically, the invention relates to a method of preparing an expression vector containing linked exon sequences comprising amplifying exon sequences by PCR as one or more fragments from a giant fungal gene containing an intron, and linking the fragments together with a restriction enzyme-treated vector using the gap repair cloning method; a method of preparing an expression vector containing a full-length cDNA sequence by synthesizing and linking cDNA fragments from a fungal giant gene; a transformant having introduced therein an expression vector prepared by the method; a protein produced by the transformant; and a method of preparing a compound produced by the protein using the expression vector.
机译:本发明涉及从真核基因中去除基因中包含的内含子,并仅连接外显子序列以制备包含连接的序列的表达载体的方法。具体地,本发明涉及一种制备包含连接的外显子序列的表达载体的方法,该方法包括通过PCR从包含内含子的巨型真菌基因中扩增作为一个或多个片段的外显子序列,并将所述片段与使用限制性酶处理的载体连接在一起。缺口修复克隆方法;一种通过合成和连接来自真菌巨基因的cDNA片段来制备含有全长cDNA序列的表达载体的方法。导入了通过该方法制备的表达载体的转化体。转化体产生的蛋白质;以及使用表达载体制备由蛋白质产生的化合物的方法。

著录项

  • 公开/公告号US8980580B2

    专利类型

  • 公开/公告日2015-03-17

    原文格式PDF

  • 申请/专利权人 KENJI WATANABE;HISAO MORIYA;

    申请/专利号US201113816826

  • 发明设计人 KENJI WATANABE;HISAO MORIYA;

    申请日2011-08-11

  • 分类号C12Q1/68;C12P19/34;

  • 国家 US

  • 入库时间 2022-08-21 15:20:25

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