盐藻（Dunaliella viridis）细菌人工染色体文库的构建和鉴定

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.

Construction and Characterization of a Dunaliella viridis Bacterial Artificial Chromosome Library

Dunaliella 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.