数字农科院2.0

草酸青霉菌11的cDNA文库构建及其溶磷相关基因的筛选

文献类型： 中文期刊

作者： 唐超西;龚明波;李顺鹏;朱昌雄

作者机构：

关键词： 草酸青霉菌;eDNA文库;溶磷相关基因;有机酸;Penicillium oxalicum; cDNA library; phosphate-solubilizing related gene; organic acid

期刊名称：中国农业科学

ISSN： 0578-1752

年卷期： 2012 年 45 卷 018 期

页码： 3792-3800

收录情况： 北大核心 ; CSCD

摘要： 【目的】构建草酸青霉菌11的cDNA文库,筛选溶磷相关基因。【方法】利用SMART技术构建草酸青霉菌11的初级cDNA文库,通过难溶磷培养基筛选具有溶磷能力的转化子,测序并进行生物信息学分析。在难溶磷液体培养基中,进行转化子对溶液pH值、可溶磷含量的影响和产有机酸试验。【结果】成功构建了草酸青霉菌11的初级cDNA文库,其库容量约为5.29×10^6cfu·mL-1,重组率为99%;利用难溶磷固体培养基筛选,得到具有溶磷圈的转化子48个,其中转化子I一4的cDNA序列全长536bp,为一个新的序列,基因编码氨基酸残基序列长129n.t。转化子EcoliHST081-4在液体难溶磷培养基中培养,提高了有机酸的表达量,并增加了有机酸的种类,在培养i2h后,开始产生乙酸,24h后,溶液中产生乳酸、苹果酸和α-酮戊二酸,培养36h,溶液pH值由6.32降到3.69,可溶磷含量达到0.1076mg·mL-1。【结论】从草酸青霉II中筛选到一个溶磷相关基因pStI.

分类号： Q78

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