Obtaining and analysis of flanking sequences from T-DNA transformants of Arabidopsis

被引：65

作者：

Qin, GJ ^{[1
]}

Kang, DM ^{[1
]}

Dong, YY ^{[1
]}

Shen, YP ^{[1
]}

Zhang, L ^{[1
]}

Deng, XH ^{[1
]}

Zhang, Y ^{[1
]}

Li, S ^{[1
]}

Chen, N ^{[1
]}

Niu, WR ^{[1
]}

Chen, C ^{[1
]}

Liu, PC ^{[1
]}

Chen, HD ^{[1
]}

Li, JG ^{[1
]}

Ren, YF ^{[1
]}

Gu, HY ^{[1
]}

Deng, XW ^{[1
]}

Qu, LJ ^{[1
]}

Chen, ZL ^{[1
]}

机构：

[1] Peking Univ, Coll Life Sci, Natl Lab Prot Engn & Genet Engn, Peking Yale Joint Ctr Plant Mol Genet & Agrobiote, Beijing 100871, Peoples R China

来源：

PLANT SCIENCE | 2003年 / 165卷 / 05期

关键词：

T-DNA; Arabidopsis mutant collection; flanking sequence; location; distribution;

D O I：

10.1016/S0168-9452(03)00218-8

中图分类号：

Q5 [生物化学]; Q7 [分子生物学];

学科分类号：

071010 ; 081704 ;

摘要：

Large collections of insertional Arabidopsis lines are valuable for research on functional genomics. Using the activation tagging vector pSKI015, more than 45000 T-DNA insertion lines were generated by Agrobacterium-mediated floral-dip transformation protocol. 2304 insertion lines were analyzed and 1502 items of plant sequences flanking the T-DNA insertion sites were obtained by a modified thermal asymmetric interlaced PCR (TAIL-PCR) protocol. These sequences were searched against Genebank database using BLAST and 1194 insertion sites were determined according to the sequences matching to the Arabidopsis genome sequence. The insertion sites were distributed on all the five Arabidopsis chromosomes and the interfered genes were classified into 14 function categories. Analysis on 1194 items of 100-bp sequences surrounding T-DNA insertion sites showed that 27 and 31% GC contents were likely to favor the T-DNA integration. Sixty-eight items of these 100-bp sequences having more than two insertions were chosen to look for motifs in favor of T-DNA integration. The results showed that "ATNTT" (N represents A/T/C/G) and the polyT and polyA motifs probably play a role in the T-DNA integration event. (C) 2003 Elsevier Ireland Ltd. All rights reserved.

引用

页码：941 / 949

页数：9

共 25 条

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