| 1 | 高覆盖率水稻BAC库的构建及抗病基因相关克隆的筛选显示文摘利用含Xa4、xa5和xa13 3个水稻白叶枯病抗性基因的累加系IRBB56构建了一个水稻细菌人工染色体文库。该文库包含55 296个克隆,平均插入片段为132kb。按水稻基因组为450Mb计,该文库覆盖14倍基因组,筛选出任一水稻基因或序列的概率为99.99%。用均匀分布的3个叶绿体基因和4个线粒体基因克隆作探针筛选文库,结果显示该文库中含细胞器基因组DNA同源序列的克隆数小于1%。用分布于水稻3条不同染色体、分别与Xa4、xa5和xa13连锁的DNA标记筛选文库,分别检测出11~106个阳性克隆,为克隆这些基因打下了基础。 该文库对水稻基因组的高度覆盖率和较大的插入片段,非常适合于物理作图和基因的分离和克隆。 | 王文明 江光怀 王世全 朱立煌 翟文学 | 2001 | Acta Genetica Sinica2001，28，2： | 25 |
| 2 | Construction and characterization of the transformation-competent artificial chromosome (TAC) libraries of Leymus multicaulis显示文摘Transformation-competent artificial chromosome system is able to clone and transfer genes efficiently in plants.In order to clone genes highly tolerant to barley yellow dwarf virus(BYDV),Aphids,drought and salt from Leymus multicaulis,the two TAC genomic libraries I and II were constructed in vector pYLTAC17 and pYLTAC747H/sacB,which contain about 165000 and 236000 recombinant clones sepa-rately.The genome coverage of the two libraries was totally estimated to be about 3―5 haploid genome equivalents,as size selection of genomic DNA fragments was approximately from 9 to 300 kb.Clones of the genomic libraries were collected as bulked pools each containing 500 clones or so,stored in twelve 96-deep-well plates and then were gridding in triplicate onto a high-density colony hybridization filter with a 3×3 pattern using a GeneTAC?G3 arraying robot after being transferred manually into three 384-well plates.Meanwhile 2501 and 2890 clones of Library in pYLTAC17 and in pYLTAC747H/sacB were stored individually in fourteen 384-well plates and then were automatically gridding in duplicate onto a high-density colony hybridization filter with a 6×6 pattern after a replication of plates.Nineteen positive clones were detected by using the probe glutahione reductase gene of L.multicaulis.TAC libraries constructed here can be used to isolate genomic clones containing target genes,and to carry out genome walking for positional cloning.Once the target TAC clones were isolated,they could be immediately transferred into plant genomes with the Agrobacterium system. | XU YueYu 1,2,3,ZHOU YuLei 4,5 ,SONG LinLin 6 ,ZHANG Yan 3 &ZHAO MaoLin 1 1 Beijing Agro-Biotechnology Research Center,Beijing Academy of Agriculture and Forestry Science,Beijing 100097,China 2 Department of Chemistry and Chemical Engineering,Hunan Institute of Engineering,Xiangtan 411104,China 3 College of Life Science,Capital Normal University,Beijing 100037,China 4 College of Life Science,Zhongkai University of Agriculture and Technology,Guangzhou 510225,China 5 College of Grassland Science,Gansu Agricultural University,Lanzhou 730070,China 6 Department of Biology,Henan Institute of Science and Technology,Xinxiang 453003,China | 2008 | Science China(Life Sciences)2008，51，7： | 1 |
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