Systematic reverse genetic screening of T-DNA tagged genes in rice for functional genomic analyses: MADS-box genes as a test case.
We have generated 47 DNA pools and 235 subpools from 21,049 T-DNA insertion lines of rice. DNA pools of 500-1,000 lines were adequate for screening a T-DNA insertion within a 2-kb region. To examine the efficacy of the DNA pools, we selected MADS-box genes, which play an important role in controlling various aspects of plant development. A total of 34 MIKC-type MADS-box genes have now been identified from rice sequence databases. Our PCR screening for T-DNA insertions within 12 MADS-box genes resulted in the identification of five insertions in four different genes. These DNA pools will be valuable when isolating T-DNA insertional mutants in various rice genes. The DNA pool screening service and the mutant seeds are available upon request to genean@postech.ac.kr.
Lee
Shinyoung
S
National Research Laboratory of Plant Functional Genomics, Division of Molecular and Life Sciences, Pohang University of Science and Technology (POSTECH), Pohang 790-784, Republic of Korea.
Kim
Joonyul
J
Son
Jun-Seock
JS
Nam
Jongmin
J
Jeong
Dong-Hoon
DH
Lee
Keunsub
K
Jang
Seonghoe
S
Yoo
Jihye
J
Lee
Jinwon
J
Lee
Dong-Yeon
DY
Kang
Hong-Gyu
HG
An
Gynheung
G
D004269:DNA, Bacterial / Q000737:chemistry / Q000235:genetics; D018745:Genome, Plant; D026161:MADS Domain Proteins / Q000235:genetics*; D009154:Mutation; D012275:Oryza / Q000737:chemistry / Q000235:genetics*; D010802:Phylogeny; D010940:Plant Proteins / Q000235:genetics; D020133:Reverse Transcriptase Polymerase Chain Reaction
D004269:DNA, Bacterial; D026161:MADS Domain Proteins; D010940:Plant Proteins; C036483:T-DNA