乌鲁木齐10号冷泉细菌群落密度实时荧光定量PCR检测体系的建立  

The Preliminary Establishment of for Detecting the Community Bacterial Density from No.10 Spring in Urumqi by the Real-Time PCR System

作　　者：唐凤[1,2] 史应武[2] 李萍[1] 罗娇[3] 晁群芳[1] 

机构地区：[1]新疆大学生命科学与技术学院,乌鲁木齐830046 [2]新疆农业科学院微生物应用研究所,乌鲁木齐830091 [3]石河子大学生命科学学院,新疆石河子832003

出　　处：《新疆农业科学》2015年第10期1872-1877,共6页Xinjiang Agricultural Sciences

基　　金：新疆维吾尔自治区高新技术发展项目(201216145)~~

摘　　要：【目的】初步建立乌鲁木齐10号冷泉泉水细菌群落的定量分析体系。【方法】选用地震前与地震后的乌鲁木齐10号冷泉水样,以及无震时期的水样做为三种处理方式,利用构建克隆子的方法制作标准样品,并对实时荧光定量PCR标准曲线的线性(R^2)、斜率(S)、扩增效率(E)等相关参数进行优化。【结果】标准曲线的线性R^2=0.993、以及斜率S=-3.415,E=1.963均基本符合参数要求。震前冷泉水体细菌16 S rDNA片段拷贝数达到1.893×10~5 cop/mL,震后冷泉水体细菌16 S rDNA片段拷贝数达到1.43×10~6 cop/mL无震时期水体细菌16 S rDNA片段拷贝数达到9.931×10~6 cop/mL。【结论】建立的实时荧光定量PCR检测体系符合参数要求,并且可以灵敏地、快速地定量未知模板的乌鲁木齐10号泉细菌数。[ Objective ] Preliminary establishment of the system of quantitative analysis for detecting the bacterial community abundance from No. 10 Spring in Urumqi. [ Method] Choosing pie - and post - earthquake of Urumqi No. 10 Spring water, and no vibration period of water as three kinds of processing methods. Using the molecular cloning method, the standard substance of real - time PCR was structured, and the relevant parameter was optimized including the linearity （R2 ）, amplification efficiency （E） and slope （S） of the standard curve. [ Result ] The standard curve parameter was in line with the basic parameter requirements, which were R2 = 0. 993 ,S = - 3. 415 and E = 1. 963. The pre - and post - earthquake and non earthquake period of the No. 10 Spring water bacteria 16 S rDNA fragment of copy number reached respectively 2.767 ×10^4 cop/mL, 3. 332×10^5 cop/mL, 9. 931×10^6 cop/mL. [ Conclusion] Real time fluorescence quantitative PCR detection system has been initially established which was consistent with parameter requirements, and which was sensitive and rapid to determine the bacterial number of the unknown template of Urumqi No. 10 spring.

关 键 词：乌鲁木齐10号冷泉 实时荧光定量PCR 绝对定量 

分 类 号：S182[农业科学—农业基础科学]