贵州医科大学学报

2008, No.130(01) 1-4

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贵阳腐霉几丁质酶的活性及基因片段克隆与序列
Determination of Chitinase Activity of Pythium guiyangense Su and the Cloning and Analysis of A Fragment of the Chitinase Gene

洪焱;苏晓庆;
HONG Yan, SU Xiaoqing(Department of Biology, Guiyang Medical College, Guiyang 550004, Guizhou, China)

摘要(Abstract):

目的:研究贵阳腐霉(Pythium guiyangense Su)几丁质酶的活性及其基因片段的克隆与序列分析。方法:(1)采用3,5-二硝基水杨酸比色法测定贵阳腐霉分泌的几丁质酶活性;(2)根据NCBI Protein数据库中多种生物的几丁质酶氨基酸序列的保守区设计引物,以贵阳腐霉基因组DNA为模版,通过PCR扩增获得特异性DNA片段;(3)将该片段克隆并进行测序分析。结果:贵阳腐霉经诱导后可以分泌几丁质酶。诱导24h是该酶分泌高峰期。克隆到一207bp的DNA片段,其第182~207碱基序列与狗(Canis familiaris)的几丁质酶基因的部分序列相同。结论:本实验证明贵阳腐霉核DNA含有几丁质酶基因,在几丁质类似物的诱导下,该菌可以分泌几丁质酶。所克隆的DNA片段可能是几丁质酶基因的部分序列。
Objective: To test the chitinase activities produced by Pythium guiyangense Su, and to clone and sequence a fragment of the chitinase gene. Methods: 3,5-dinitrosalicylic acid chromatometry was used to test chitinase activities; DNA fragment was obtained with PCR technique using primers designed following conservative areas of chitinases of other organisms and sequenced. Results: It was demonstrated that P. guiyangens can produce chitinase after the induction of chitin-like substances, and the peak time of secretion was at about 24 hours after the induction. A 207 bp DNA fragment was obtained which was partly identical with a chitinase gene of Canis familiaris. Conlusions: P. guiyangense has chitinase gene, and can produce chitinase under induction. The obtained DNA fragment might be a part of chtinase gene of P. guiyangense, which needs further study to confirm.

关键词(KeyWords): 真菌;灭蚊;聚合酶链反应;贵阳腐霉;几丁质酶;几丁质酶活性
fungi; mosquito control; polymerase chain reaction; Pythium guiyangense Su; chitinase; chitinase activity

Abstract:

Keywords:

基金项目(Foundation): 国家自然科学基金(批准号30160003);; 贵州省国际科技合作重点项目[合同号黔科合外字(2005)400118号]

作者(Author): 洪焱;苏晓庆;
HONG Yan, SU Xiaoqing(Department of Biology, Guiyang Medical College, Guiyang 550004, Guizhou, China)

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DOI: 10.19367/j.cnki.1000-2707.2008.01.002

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