大豆疫霉多聚半乳糖醛酸酶pspg1基因毕赤酵母表达载体的构建Construction of Expression Vector of Polygalacturonase Gene pspg1 from Phytophthora sojae
孙文秀;
摘要(Abstract)：
[目的]构建大豆疫霉多聚半乳糖醛酸酶pspg1基因毕赤酵母表达载体。[方法]利用PCR方法从大豆疫霉菌cDNA中扩增多聚半乳糖醛酸酶pspg1基因,用限制性内切酶EcoRⅠ和NotⅠ分别酶切此基因和毕赤酵母表达载体pPIC9K,然后用T4连接酶将目的基因克隆到pPIC9K载体中构建此载体。[结果]扩增的大豆疫霉菌pspg1基因成熟肽片段大小为1 250 bp。用基因特异性引物扩增阳性重组子,可得到大小为1 200 bp的片段。而用载体特异性引物扩增阳性重组子,可得到大小为1 200和1 700 bp两条带,多出来的400 bp恰好符合载体部分的大小。将此特异性片段PCR扩增回收,序列测定发现大豆疫霉pspg1基因已经成功地插入到表达载体中。[结论]该研究为进一步研究大豆疫霉菌pspg1基因在毕赤酵母中高效表达奠定了基础。
关键词(KeyWords)： 大豆疫霉菌;多聚半乳糖醛酸酶;毕赤酵母表达载体;构建
基金项目(Foundation): 长江大学博士创新基金项目
作者(Author): 孙文秀;
Email:
DOI: 10.13989/j.cnki.0517-6611.2009.19.058
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