Fluorescence in situ hybridization analysis of alien genes in Agrobacterium-mediated Cry1A(b)-transformed rice.
The transgene in Agrobacterium-mediated Cry1A(b)-transgenic rice plants has been detected and its chromosomal location determined by fluorescence in situ hybridization (FISH). Eight of the nine transgenic lines tested showed hybridization signals. Signals were located on regions of the chromosome in which fraction length (FL) values varied from 26.2 (near the centromere) to 95.2 (distal regions). No signal was found on regions where the fraction length was less than 26.2, while six of the nine signals detected were located on regions with FL values of 75.3 or over. This demonstrates that Agrobacterium-mediated genes can integrate into multiple sites distributed in different parts of the chromosome, but that distal regions are the preferred sites and regions near the centromeres are colder for T-DNA integration. The donor DNA of the transformation was divided into two parts, labelled separately as probes for two-colour FISH. Results show that the transformed DNA sequences remained linked in the recipient genome. The relationship between integration position and transgene silencing, known as the 'position effect', is discussed.
Key Lab of MOE for Plant Developmental Biology, Wuhan University, PR China.
Wei-Wei
WW
Jin
Zong-Yun
ZY
Li
Qing
Q
Fang
Illimar
I
Altosaar
Li-Hua
LH
Liu
Yun-Chun
YC
Song
D056931:Cryptochromes; D015342:DNA Probes; D029721:Drosophila Proteins; D005136:Eye Proteins; D005420:Flavoproteins / Q000235:genetics*; D005798:Genes, Bacterial; D017404:In Situ Hybridization, Fluorescence; D012275:Oryza / Q000235:genetics*; D017956:Photoreceptor Cells, Invertebrate; D012333:RNA, Messenger / Q000235:genetics; D043562:Receptors, G-Protein-Coupled; D014170:Transformation, Genetic
D056931:Cryptochromes; D015342:DNA Probes; D029721:Drosophila Proteins; D005136:Eye Proteins; D005420:Flavoproteins; D012333:RNA, Messenger; D043562:Receptors, G-Protein-Coupled; C119828:cry protein, Drosophila