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盐藻(Dunaliella viridis)细菌人工染色体文库的构建和鉴定 被引量：3

Construction and Characterization of a Dunaliella viridis Bacterial Artificial Chromosome Library
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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55kb,以单克隆形式保藏在96块96孔板中,并建立了四维PCR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BAC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. DunalieUa viridis has outstanding ability to adapt to hyper-salinity, and is a model system for plant salt tolerance mechanism study. We constructed a bacterial artificial chromosome library of Dunaliella viridis, using vector pCC1BAC. The library consisted of 9 216 clones on 96 96-well plates. The average insertion size was about 55 kb. A four-dimensional PCR screening system was developed, by which single positive clone could be obtained with 4 rounds of PCR. Based 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.
作者 孟祥宗 杨晓武 管祯玮 张男 许政暟 宋任涛
机构地区 上海大学生命科学学院
出处 《上海大学学报（自然科学版）》 CAS CSCD 北大核心 2007年第1期88-93,共6页 Journal of Shanghai University:Natural Science Edition
基金 国家自然科学基金资助项目(3047111930470857)
关键词 盐藻 细菌人工染色体文库 四维PCR筛选体系 Dunaliella viridis bacterial artificial chromosome （BAC） library four-dimensional PCR （4DPCR） screening system
分类号 Q785 [生物学—分子生物学]
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参考文献13

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二级参考文献28

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共引文献14

同被引文献31

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  • 3万国福,谷绒,唐会英,车振明.超声波处理在果胶提取工艺中的应用[J].食品研究与开发,2006,27(7):115-117. 被引量：30
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引证文献3

二级引证文献1

上海大学学报（自然科学版）
2007年 第1期

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