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OALib Journal期刊
ISSN: 2333-9721
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蔗糖富集环境土壤微生物宏基因组分析及蔗糖水解相关酶基因克隆
DOI: 10.3724/SP.J.1145.2010.00403, PP. 403-407
Keywords: 宏基因组,环境dna,dna文库,细菌多样性统计技术,蔗糖水解酶
Abstract:
蔗糖水解酶是蔗糖转化生成生物质能源的关键酶，且还具有重要的转糖苷功能.针对蔗糖富集的土壤环境，利用未培养的宏基因组技术对蔗糖水解相关的酶基因进行克隆.首先使用微生态分子技术对蔗糖富集的土壤样品进行分析，在可信区间为95%的情况下，样品覆盖率为20%（c指数为0.2），speciesrichness指数为235.0，shannonindex为5.2889，说明这个蔗糖富集样品中的微生物来源具有广泛性.然后使用宏基因组技术构建这个土壤样品中微生物的dna文库，成功构建一个包含约100000个克隆的大片段dnafosmid文库.对文库中的fosmid质粒进行随机测序，发现质粒的外源dna与已报道的dna都没有同源性，文库所克隆的dna都来源于仍没有被研究的微生物.使用蔗糖作为唯一碳源对文库进行筛选,获得了能水解蔗糖的克隆.在蔗糖水解能力最强的两个克隆中所包含的蔗糖水解酶与genbank数据库中已知蔗糖酶的相似性分别为38%和68%.图2表2参27
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