| 摘要: |
| 研究优化了樟疫霉菌多聚半乳糖醛酸酶Pcpg16和Pcpg17基因克隆的PCR条件，并对其进行了克隆、测序和遗传转化研究。克隆获得了2个基因，其大小均为996 bp；通过构建表达载体和遗传转化获得了2个基因的转基因菌系；这2个基因均能指导合成相应的PG酶。与对照AnpgⅠ相比，转化Pcpg17基因的酵母菌所分泌的多聚半乳糖醛酸酶（PG）活性较弱，而Pcpg16基因指导合成的PG酶无活性。Western blotting表明，所克隆的2个基因指导合成的PG酶均有不同程度的糖基化。 |
| 关键词: 疫病病原菌 Pcpg基因 基因克隆与测序 基因表达 PG活性 |
| DOI： |
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| 基金项目:陕西省农业分子生物学重点实验室基金项目(2002-2) |
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| Studies on cloning,sequencing and Saccharomyces cerevisiae expression of Pcpg (Phytophthora cinnamomi polygalacturonase) 16 and Pcpg17 |
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| Abstract: |
| The components of the polymerase chain reaction and reaction parameters for cloning Pcpg (Phytophthora cinnamomi polygalacturonase) 16 and Pcpg17 genes were optimized.The genes were sequenced,their expression vectors were determined and the genes were transformed to a yeast line,W303-1B.Then the PG activity of transgenic lines was investigated.The genes were cloned and both were 996 bp.Both Pcpg16 and Pcpg17 transgenic yeast lines were obtained，and although the two genes could synthesize the PGs,the yeast line of Pcpg16 gene did not show PG activity.The PG activity of the transgenic line of Pcpg17 gene was weaker than that of the control,Anpg (Aspergillus niger polygalacturonase)Ⅰ.Western blotting results showed that there were different degrees of glycosylation of PGs encoded by Pcpg16 and Pcpg17 genes. |
| Key words: Phytophthora cinnamomi Pcpg gene gene cloning and sequencing gene expression PG activity |
