摘要： 采用农杆菌介导转化方法，将携带有GUS基因1301质粒转入日本晴水稻品种中，建立水稻T-DNA插入群体5200个。研究表明:通过PCR和SouthernBlot分析证明了再生植株为转基因植株；通过改进热不对称交错PCR(TAIL-PCR)方法，已成功地分离并测定了437个株系的侧翼序列；通过序列同源性分析表明，有147个序列只包含有载体序列，有214个序列中包含有与T-DNA右边界相邻的水稻基因组序列；通过水稻边界序列与网上水稻数据库中基因组序列比对，将3个突变体的T-DNA插入位点定位于水稻特定染色体上。
关键词:
T-DNA,
插入突变,
转基因水稻,
TAIL-PCR
Abstract: Abstract:In this study the method of Agrobacterium-mediated transformation was used to transform GUS genecarried by plasmid 1301 into Nipponbare and 5200 rice TINA insertion population had been established. Studiesshowed that transgenic plants had been proved to be transformed by methods of PCR and Southern Blot and 437 flan-king sequence had been isolated by improving method of TAILCR. By sequence homology analysis showed that therewere 147 sequences containing only the vector sequence，and 214 sequences containing TINA right border with theneighboring ricesequences and public databases in the rice genomesequence，threegenome sequence. By comparison of the bordermutants TINA insertion sites had been locatedin a particular rice chromosome. These results willprovide important information for functional genomics research.
Key words:
TINA,
Insertional mutaenesis,
Transenic ICeTAILCR
中图分类号:
魏进招, 陈秋玲, 裴忠有, 孙守钧. 水稻T-DNA插入群体的建立及侧临序列的获得与分析[J]. 华北农学报, 2011, 26(1): 31-36. doi: 10.7668/hbnxb.2011.01.007.
WEI Jin-zhao, CHEN Qiu-ling, PEI Zhong-you, SUN Shou-jun. Construction of Rice TINA Insertion Population and the Acquisitionand Analysis of Flanking Sequence[J]. ACTA AGRICULTURAE BOREALI-SINICA, 2011, 26(1): 31-36. doi: 10.7668/hbnxb.2011.01.007.