首页|MassARRAY单核苷酸多态性MALDI-TOF质谱基因分型法检测未成熟冬虫夏草子座中多个冬虫夏草菌突变基因型

MassARRAY单核苷酸多态性MALDI-TOF质谱基因分型法检测未成熟冬虫夏草子座中多个冬虫夏草菌突变基因型

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目的:检测未成熟冬虫夏草子座中同时存在的多种突变基因型冬虫夏草菌.方法:采用MassARRAY单核苷酸多态性(single nucleotide polymorphism,SNP)质谱基因分型法,根据冬虫夏草菌基因内转录间隔区(ITS)序列的大量散在点突变,设计8个SNP延伸引物,利用质谱测定各个SNP位点的单核苷酸延伸反应,检测未成熟冬虫夏草子座中与GC偏倚基因型(gb#AB067721)冬虫夏草菌相关的突变基因型.结果:GC偏倚基因型和两个AT偏倚基因型同时存在于未成熟天然冬虫夏草子座中.SNP基因分型法还检测到另外至少两个未知序列的突变基因型.结论:已知的3个冬虫夏草菌基因型的共存提示在冬虫夏草的子座萌发阶段和成熟过程中可能存在DNA转换点突变;另外两个未知序列的基因型的存在提示冬虫夏草中可能存在DNA颠换点突变.
Detection of multiple Ophiocordyceps sinensis mutants in premature stroma of Cordyceps sinensis by MassARRAY SNP MALDI-TOF mass spectrum genotyping
Objective : To examine the mutants of Ophiocordyceps sinensis ( Os) in the stroma of premature Cordyceps sinensis ( Cs). Methods: Used MassARRAY single nucleotide polymorphism ( SNP) MALDI-TOF mass spectrum genotyping, designed eight SNP extension primers on the basis of the scattered, multiple point mutations of known Os mutants within their internal transcribed spacer (ITS) segments, and examined the Os mutant genotypes relating to the GC-biased Os genotype ( gb #AB067721 ) in premature Cs stroma. Results : The two AT-biased genotypes and the GC-biased Os were simultaneously detected in premature Cs stroma. SNP genotyping also detected at least two other Os genotypes of unknown sequences. Conclusion: Coexistence of the three known Os genotypes indicates the existence of possible transition point mutations within Os genes during germination and early maturation of Cs. Simultaneous detection of at least two unknown genotypes coexisting with those known mutants possibly evidences the transversion mutations within Os genes.

高凌、李晓红、赵建晴、陆继红、朱佳石、ZHU Jia-shi

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美国华茂公司北京临床药理研究中心,北京,100088

博奥生物有限公司,北京,102206

Pharmanex Research Institute,Provo UT 84601,USA

新疆石河子大学药学院,石河子,832000

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冬虫夏草 基因型 突变 多态性 单核苷酸

华茂冬虫夏草研究基金博奥生物有限公司科研基金

2011

北京大学学报(医学版)
北京大学

北京大学学报(医学版)

CSTPCDCSCD北大核心
影响因子:1.681
ISSN:1671-167X
年,卷(期):2011.43(2)
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