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红树林土壤细菌群落16S rDNA V3片段PCR产物的DGGE分析
引用本文:王岳坤,洪葵.红树林土壤细菌群落16S rDNA V3片段PCR产物的DGGE分析[J].微生物学报,2005,45(2):201-204.
作者姓名:王岳坤  洪葵
作者单位:1. 中国热带农业科学院热带生物技术研究所,海口,571101;中国热带农业科学院橡胶研究所,儋州,571737
2. 中国热带农业科学院热带生物技术研究所,海口,571101
基金项目:“8 63”海洋生物技术青年基金 (2 0 0 2AA62 814 0 )~~
摘    要:从土壤中抽提微生物总DNA ,直接扩增 16SrDNAV3片段 ,应用变性梯度凝胶电泳 (DGGE)和分子克隆技术分析 16SrDNAV3片段的多态性 ,发现地域因素和红树品种都是影响土壤细菌群落结构的因素。通过对杯萼海桑土壤 16SrDNAV3片段PCR产物两个DGGE条带进行分子克隆、序列测定和Blast分析 ,发现每个DGGE条带包含着许多不同的 16SrDNAV3片段 ,并且其中多数为NCBI未收录的序列。这表明DGGE和克隆技术相结合的方法是研究土壤微生物群落结构的一种可行方法。

关 键 词:红树林土壤  16SrDNA  DGGE
文章编号:0001-6209(2005)02-0201-04
修稿时间:2004年8月12日

Mangrove soil community analysis using DGGE of 16S rDNA V3 fragment polymerase chain reaction products
WANG Yue-kun,HONG Kui.Mangrove soil community analysis using DGGE of 16S rDNA V3 fragment polymerase chain reaction products[J].Acta Microbiologica Sinica,2005,45(2):201-204.
Authors:WANG Yue-kun  HONG Kui
Institution:Institute of Tropical Biological Sciences and Biotechnology, Chinese Academy of Tropical Agrocultural Science, Haikou 571101, China. xwangyk@126.com
Abstract:The 16S rDNA V3 fragment polymerase chain reaction products amplified from mangrove soil-DNA extracts were analyzed by denaturing gradient gel electrophoresis (DGGE) followed by clone technology. Regional factors and mangrove species were found to influence the soil microbe community. Two DNA slices separated on denaturing gradient gel from Sonneratia alba soil sample were recovered and the recovered DNA were used to establish two clone libraries. Some clones were sequenced and each sequence was compared with all nucleotide sequences in GenBank database. Most cloned sequences were found not in GenBank database. These results suggest that DGGE followed by clone technique is a practicable protocol to research the complex community of soil microbe.
Keywords:Mangrove soil  16S rDNA  DGGE
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