A simple method to transfer, integrate and study expression of foreign genes, such as chicken ovalbumin and alpha-actin in plant tumors.
A simple method for inserting foreign genes into the T-region of Agrobacterium Ti-plasmids is described. A modified cosmid (pHC 79) was introduced into a predetermined site of the T-region of pTi C58. An Agrobacterium strain harboring this modified Ti-plasmid was used as an acceptor strain into which genes, cloned in pBR322, can be introduced by mobilization from Escherichia coli. pBR322-derived plasmids cannot replicate in Agrobacterium, but can be maintained by integration into the T-region of the modified Ti-plasmids by homologous recombination. This method was used to introduce the genes for ovalbumin and alpha-actin from chicken into tobacco tumors. Southern blotting and re-isolation of the inserted genes by reverse cloning showed that the animal DNA was transferred and integrated into the plant genome without rearrangements. The alpha-actin gene is not transcribed in plant tumors, whereas transcription of the ovalbumin gene was observed, however the initiation point of transcription was different from the one used in the chicken oviduct. The RNA transcribed from the chicken ovalbumin gene is polyadenylated and ranges in size between 2 and 7 kb.
28 references, click to expand
-
Site-specific insertion of genes into T-DNA of the Agrobacterium tumor-inducing plasmid: an approach to genetic engineering of higher plant cells.J Mol Appl Genet. 1981;1(1):39-49 PMID: 6955419
-
Labeling deoxyribonucleic acid to high specific activity in vitro by nick translation with DNA polymerase I.J Mol Biol. 1977 Jun 15;113(1):237-51 PMID: 881736
-
Rapid mapping of transposon insertion and deletion mutations in the large Ti-plasmids of Agrobacterium tumefaciens.Nucleic Acids Res. 1979 Dec 11;7(7):1837-49 PMID: 231764
-
The functional organization of the nopaline A. tumefaciens plasmid pTiC58.Plasmid. 1980 Mar;3(2):212-30 PMID: 6100894
-
A rapid micro scale method for the detection of lysopine and nopaline dehydrogenase activities.Biochim Biophys Acta. 1978 Dec 8;527(2):497-500 PMID: 31918
-
Molecular cloning of overlapping segments of the nopaline Ti-plasmid pTiC58 as a means to restriction endonuclease mapping.Plasmid. 1980 Mar;3(2):193-211 PMID: 6100893
-
A small cosmid for efficient cloning of large DNA fragments.Gene. 1980 Nov;11(3-4):291-8 PMID: 6260575
-
Regeneration of intact tobacco plants containing full length copies of genetically engineered T-DNA, and transmission of T-DNA to R1 progeny.Cell. 1983 Apr;32(4):1033-43 PMID: 6301678
-
Rapid isolation of high molecular weight plant DNA.Nucleic Acids Res. 1980 Oct 10;8(19):4321-5 PMID: 7433111
-
The ovalbumin gene: cloning and molecular organization of the entire natural gene.Proc Natl Acad Sci U S A. 1979 May;76(5):2253-7 PMID: 287066
-
Ovalbumin is synthesized in mouse cells transformed with the natural chicken ovalbumin gene.Proc Natl Acad Sci U S A. 1980 Jan;77(1):244-8 PMID: 6928616
-
Plasmids specifying plant hyperplasias.Annu Rev Microbiol. 1981;35:531-65 PMID: 7027906
-
Plasmid vehicles for direct cloning of Escherichia coli promoters.J Bacteriol. 1979 Nov;140(2):400-7 PMID: 387734
-
Intergeneric transfer and exchange recombination of restriction fragments cloned in pBR322: a novel strategy for the reversed genetics of the Ti plasmids of Agrobacterium tumefaciens.EMBO J. 1983;2(3):411-7 PMID: 11894957
-
UV-induction of chalcone synthase mRNA in cell suspension cultures of Petroselinum hortense.Proc Natl Acad Sci U S A. 1983 May;80(9):2591-3 PMID: 16593307
-
The TL-DNA in octopine crown-gall tumours codes for seven well-defined polyadenylated transcripts.EMBO J. 1982;1(1):139-46 PMID: 16453403
-
Detection of specific sequences among DNA fragments separated by gel electrophoresis.J Mol Biol. 1975 Nov 5;98(3):503-17 PMID: 1195397
-
Expression of the Herpes simplex virus thymidine kinase gene in Saccharomyces cerevisiae.Mol Gen Genet. 1981;184(3):386-93 PMID: 6278252
-
DNA sequence preference of the progesterone receptor.Proc Natl Acad Sci U S A. 1983 Jan;80(1):16-20 PMID: 6571991
-
Large plasmid in Agrobacterium tumefaciens essential for crown gall-inducing ability.Nature. 1974 Nov 8;252(5479):169-70 PMID: 4419109
-
A rapid alkaline extraction procedure for screening recombinant plasmid DNA.Nucleic Acids Res. 1979 Nov 24;7(6):1513-23 PMID: 388356
-
Construction and characterization of new cloning vehicles. II. A multipurpose cloning system.Gene. 1977;2(2):95-113 PMID: 344137
-
Plasmid ColE1 conjugal mobility: the nature of bom, a region required in cis for transfer.Mol Gen Genet. 1982;185(2):344-51 PMID: 7045589
-
Site-specific mutagenesis of Agrobacterium Ti plasmids and transfer of genes to plant cells.J Mol Appl Genet. 1981;1(2):149-64 PMID: 7050279
-
Genetic Identification of functions of TL-DNA transcripts in octopine crown galls.EMBO J. 1982;1(1):147-52 PMID: 16453404
-
Evidence that deoxyribonucleic acid sequences flanking the ovalbumin gene are not transcribed.Biochemistry. 1980 Apr 29;19(9):1755-61 PMID: 6246933
-
Interactions and DNA transfer between Agrobacterium tumefaciens, the Ti-plasmid and the plant host.Proc R Soc Lond B Biol Sci. 1979 Apr 11;204(1155):251-66 PMID: 36626
-
Genetic analysis of crown gall: fine structure map of the T-DNA by site-directed mutagenesis.Cell. 1981 Nov;27(1 Pt 2):143-53 PMID: 6276020
Contact
No. 2 Wenbo Road, Zhangqiu District, Jinan, Shandong
Qilu Normal University · Genelibs Bioinformatics Lab
750 Shunhua Rd, Jinan
2F, Bldg F, University Science Park
Tel: 0531-88819269
WeChat Official Account
Follow our WeChat subscription account for real-time updates and the latest in medical and biological research.
Business Email
E-mail: product@genelibs.com
Copyright © 2026.Genelibs Jinan Honghui Biotechnology Co., Ltd. 鲁ICP备20007159号-2
