| 摘要: |
| 研究优化了樟疫霉多聚半乳糖醛酸酶基因9（Pcpg 9）和Pcpg 10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，其大小前者1 059 bp，后者1 290 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；2个基因均能指导合成相应的PG酶蛋白，其中转化Pcpg 10基因的酵母菌所分泌的PG酶活性最强，对照AnpgⅠ的PG酶活性次之，Pcpg 9的PG酶活性最弱。Western blotting表明，所克隆的2个基因所指导合成的PG酶蛋白均有不同程度的糖基化。 |
| 关键词: 疫病病原菌 Pcpg基因 克隆 测序 遗传转化 PG活性 |
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| 基金项目:中澳合作项目；陕西省自然科学基金项目(2001SM11)；陕西省农业分子生物学重点实验室基金项目 |
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| Studies on cloning,sequencing and genetic transformation of Pcpg (Phytophthora cinnamomi polygalac-turonase) 9 and Pcpg 10 |
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| Abstract: |
| Pcpg (Phytophthora cinnamomi polygalacturonase) 9 and Pcpg 10 genes were cloned based on the studying of the PCR composition and its reaction condition of the genes.The sequence and their expression vectors of the genes were done and the genes were transformed to a yeast line of W303-1B.Then the PG activity of transgenic lines was investigated.The genes were cloned and the size of Pcpg 9 was 1 059 bp and Pcpg 10 was 1 290 bp.The transgenic yeast lines of Pcpg 9 and Pcpg 10 genes were obtained respectively.Pcpg 9,Pcpg 10 and the control,Anpg(Aspergillus niger polygalacturonase)Ⅰcould guide to synthesize the PGs.The PG activity of transgenic line of Pcpg 10 was the strongest,and that of the Pcpg 9 gene was the weakest.Western blotting results showed that there were different degree glycosylation of PGs which were encoded by Pcpg 9 and Pcpg 10 genes. |
| Key words: Phytophthora cinnamomi Pcpg gene cloning sequencing genetic transformation PG activity |
