Rice Genetics IV - IRRI books - International Rice Research Institute

Aprobe AEEprobe BpGA1633RBgus Tn p35S hphT7LBpGA2144RBIgus Tn p0sTubA1 0sTubA1–1hphT7LB1 kbBB112B118B119B121B139B1558T-DNA (R)CCCGGGGATCCGTGTTtgacAAGGGACTGACCACCCGGGGGTGTTtgacaggatatattgTGACCACCCGGGGATCCGTGACCCGGGGATCCGTGTTtgaAACCGTATAAGGGACTGACC0 bp15 bp6 bp11 bp0 bp488 bpT-DNA (L)GCTTAGACAACTTAATAACAATGGTATTCAATTTAAACACaaacAAATTGACGCTTAGACCTTAATAACACATTGCGGACttgtggtgtaaacAAATTGACATGTATTTGAAAAATAAAACT-DNARBNumberpGA1633 – – CCCGGGGA TCCGTGT T t g a c a g g at a t a t t gg c g g g t a a a c – –– – CCCG*************************************– – CCCGGGGA TC*******************************– – CCCGGGGA TCC******************************– – CCCGGGGA TCCGTGT**************************– – CCCGGGGA TCCGTGT T*************************– – CCCGGGGA TCCGTGT T t g a**********************114133pGA2144 – – GA G C T C G A T C C G T G T T t g a c a g g a t a t a t t g g c g g g t a a a c – –– – GAGC************************************ *– – GAGCTCGATCCGTGT************************* *– – GAGCTCGATCCGTGTT*************************– – GAGCTCGATCCGTGTT t ************************– – GAGCTCGATCCGTGTT t g***********************– – GAGCTCGATCCGTGTT t ga**********************114328Fig. 1. Maps of the T-DNA tagging vectors and analysis of T-DNA integration patterns. (A)Schematic diagrams of pGA1633 and pGA2144. The RB and LB in shaded boxes represent theright border and left border of T-DNA, respectively. E = EcoRI site, gus = β-glucuronidase,Tn = nopaline synthase (nos) terminator, p35S = cauliflower mosaic virus 35S promoter,hph = hygromycin phosphotransferase, T7 = transcription termination region of gene 7 of thepTiA6, I = the OsTubA4 intron 3 carrying three putative splicing acceptor and donor sites,pOsTubA4 = the promoter of the rice α-tubulin gene, and OsTubA4 = OsTubA4-1, the first intronof OsTubA4. The gus probe (probe A) and hph probe (probe B) used for DNA blot analyses areindicated. (B) Sequences at the junctions between two directly integrated T-DNA copies. Thecapitals indicate T-DNA sequences immediately next to the right (R) and left (L) bordersequences (lower case). The length of filler sequences is depicted. (C) Sequences of T-DNA atthe junction to plant DNA. The T-DNA sequences of pGA1633 and pGA2144 are shown in capitalsand the right border (RB) sequences are shown in lower case. Asterisks signify any nucleotidespresumably originated from the rice genome.Generation of T-DNA insertional tagging lines in rice 255