盐藻（Dunaliella viridis）细菌人工染色体文库的构建和鉴定

上海大学学报(自然科学版)

盐藻（Dunaliella viridis）细菌人工染色体文库的构建和鉴定

孟祥宗;杨晓武;管祯玮

1.上海大学生命科学学院，上海 200444； 2.上海市能源作物培育及利用重点实验室，上海 200444； 3.中科院上海植物生理生态研究所，上海 200032

收稿日期:2006-04-06 出版日期:2007-02-28 发布日期:2007-02-28

Construction and Characterization of a Dunaliella viridis Bacterial Artificial Chromosome Library

MENG Xiang zong;YANG Xiao wu;GUAN Zhen wei;et al

1. School of Life Sciences, Shanghai University, Shanghai 200444, China; 2. Shanghai Key Laboratory of Bio energy Crop, Shanghai 200444, China; 3. Shanghai Institute of Plant Physiology and Ecology, Chinese Academy of Scienc es, Shanghai 200032, China

Received:2006-04-06 Online:2007-02-28 Published:2007-02-28

摘要/Abstract

摘要： 盐藻具有极强的耐盐能力，是研究植物耐盐机制的模式系统.为了对其耐盐机制进行深入的研究，以pCC1BAC为载体，构建了盐藻的细菌人工染色体（BAC）文库.该文库共有9 216 个转化子，插入片段平均长度为55 kb，以单克隆形式保藏在96块96孔板中，并建立了四维P CR基因筛选体系，可以通过4轮PCR快速筛选获得阳性单克隆.根据本实验室分离到的两个盐藻基因的cDNA序列（ DvSPT2和DvTPSP ）设计引物，通过PCR从该文库中各筛到4个阳性BA C克隆，说明该文库能有效用于分离盐藻基因的基因组序列，据此推测该文库约覆盖4倍盐藻基因组序列.

关键词: 四维PCR筛选体系, 细菌人工染色体文库, 盐藻

Abstract: Dunaliella viridis has outstanding ability to adapt to hyper salinity, and is a model system for plant salt tolerance mechanism study. We constructed a bac terial artificial chromosome library of Dunaliella viridis , using vector pCC 1 BAC. The library consisted of 9 216 clones on 96 96 well plates. The average in sertion size was about 55 kb. A four dimensional PCR screening system was devel oped, by which single positive clone could be obtained with 4 rounds of PCR. Bas ed on the sequences of two cDNAs of Dunaliella viridis isolated in our lab ( DvSPT2 and DvTPSP respectively), we isolated 4 positive clones for each gene by PCR, indicating that the library had about 4 times genome coverage.

Key words: bacterial artificial chromosome (BAC) library, four dim ensional PCR (4D PCR) screening system, Dunaliella viridis

孟祥宗;杨晓武;管祯玮. 盐藻（Dunaliella viridis）细菌人工染色体文库的构建和鉴定[J]. 上海大学学报(自然科学版).

MENG Xiang zong;YANG Xiao wu;GUAN Zhen wei;et al . Construction and Characterization of a Dunaliella viridis Bacterial Artificial Chromosome Library[J]. Journal of Shanghai University（Natural Science Edition）.

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