常用农杆菌菌株名称及其Ti质粒，抗性，基因组背景。

根癌农杆菌，发根农杆菌菌种及感受态细胞现货供应Biovector NTCC Inc.

T-DNA Binary Vectors and Systems

Genotypes of some useful Agrobacterium tumefaciens strains

• LBA4404 (Ach5 pTiAch5) Sm/Sp(R) in the virulence plasmid (from Tn904); all T-DNA of pTiAch5 eliminated in pAL4404 (Hoekema et al., 1983).

• EHA101, genotype C58 pTiBo542; T-region::aph, Km(R); A281 derivative harboring pEHA101, T-DNA replaced with nptII, elimination of T-DNA boundaries uncofirmed, super-virulent (Hood et al., 1986).

• EHA105 is a Km(S) derivative of EHA101 (Hood et al., 1993).

• AGL1, genotype is AGL0 (C58 pTiBo542) recA::bla, T-region deleted Mop(+) Cb(R) [AGL0 is an EHA101 with the T-region deleted, which also deletes the aph gene] (Lazo et al., 1991).

• A281, reconstructed strain, derivative of A136 (cured C58) harboring pTiBo542, super-virulent (Hood et al., 1986).

Types

• Ach5 .......... agrocinopine, octopine type

• B6S3, A6 .... octopine type

• Bo542 ........ leucinopine, succinamopine, agropine type, vir weaker than A281

• C58, T37 .... nopaline types

• A281 ......... succinamopine, leucinopine, agrocinopine

Antibiotics

• Chloramphenicol, 100 µg/mL for strain AGL1, 10 µg/mL for LBA4404, 25 µg/mL for EHA105 and for E. coli.

• Kanamycin, 50 µg/mL for both Agrobacterium and E. coli.

• For selection of transformed rice plants we use hygromycin at 50 µg/mL and 25 µg/mL for tobacco.

Selected References

• Chen L, Zhang S, Beachy RN, Fauquet CM (1998) A protocol for consistent, large-scale production of fertile transgenic rice plants. Plant Cell Reports 18:25-31

• Christou P (1991) Production of transgenic rice (Oryza sativa L.) plants from agronomically important indica and japonica varieties via electric discharge particle acceleration of exogenous DNA into immature zygotic embryos. Biotechnology 9:957-962

• Christou P (1997) Rice transformation: bombardment. Plant Mol Biol 35:197-203.

• Deblaere R, Reynaerts A, Hofte H, Hernalsteens JP, Leemans J, and Van Montagu M (1987) Vectors for cloning in plant cells. Meth Enzymol 153:277-292

• Hajdukiewicz,P, Svab, Z, Maliga, P., (1994) The small versatile pPZP family of Agrobacterium binary vectors for plant transformation. Plant Mol Biol 25:989-994

• Hiei Y, Ohta S, Komari T, Kumashiro T (1994) Efficient transformation of rice (Oryza sativa L.) mediated by Agrobacterium and sequence analysis of the boundaries of the T-DNA.Plant J 6:271-282

• Hoekema A, Hirsch PR, Hooykaas PJJ, Schilperoort RA (1983) Binary vector strategy based on separation of vir- and T-region of the Agrobacterium tumefaciens Ti-plasmid. Nature303:179-180

• Hood EE, Helmer GL, Fraley RT, Chilton MD (1986) The hypervirulence of Agrobacterium tumefaciens A281 is encoded in a region of pTiBo542 outside of T-DNA. J Bac 168:1291-1301

• Hood EE, Gelvin SB, Melchers S, Hoekema A (1993) New Agrobacterium helper plasmids for gene transfer to plants (EHA105). Trans Res 2:208-218

• Jefferson RA, Kavanagh TA, Bevan MW (1987) GUS fusions: Beta-glucuronidase as a sensitive and versatile gene fusion marker in higher plants. EMBOJ 6:3901-3907

• Klapwijk PM, van Breukelen J, Korevaar K, Ooms G, Schilperoort RA (1980) T ransposition of Tn904 encoding streptomycin resistance into octopine Ti plasmid of Agrobacterium tumefaciens. J Bac 141:129-136

• Lazo GR, Stein PA, Ludwig RA (1991) A DNA transformation-competent Ara bidopsis genomic library in Agrobacterium. BioTechnology 9:963-967

• Ohta S, Mita S, Hattori T, Nakamura K (1990) Construction and expression in tobacco of a beta-glucuronidase (GUS) reporter gene containing an intron within the coding sequence. Plant Cell Physiol 31:805-814

• Ooms G, Hooykaas PJJ, Van Veen RJM, Van Beelen P, Regensburg-Tunk JG, Schilperoort RA (1982) Octopine Ti-plasmid deletion mutants of Agrobacterium tumefaciens with emphasis on the right side of the T-region. Plasmid 7 :15-29

• Peralta EG, Hellmiss R, Ream W (1986) Overdrive, a T-DNA transmission enhancer on the A. tumefaciens tumour-inducing plasmid. EMBO J 5:1137-1142

• Porath, J. (1992). Immobilized metal ion affinity chromatography. Protein Expre Purif 3:263-281

• Siemering KR, Golbik R, Sever R, Haseloff J (1996) Mutations that suppress the thermosensitivity of green fluorescent protein. Curr Biol 6:1653-1663

• Tanaka A, Mita S, Ohta S, Kyozuka J, Shimamoto K, Nakamura K (1990) Enhancement of foreign gene expression by a dicot intron in rice but not in tobacco is correlated with an increased level of mRNA and an efficient splicing of the intron. Nucl Acids Res 18:6767-6770

Other Resources

• List of TransBacter strains available (will be linked soon)

• Read Frequently Asked Questions about requesting CAMBIA materials

• Read Frequently Asked Questions about pCAMBIA Vectors

• GUSPlus information in the BioForge GUSPlus Project.