Kloning Gen cry dari Bacillus turingiensis var. kurstaki

https://doi.org/10.22146/jpti.9918

Nur Rahmi Ardiani(1*), Sismindari Sismindari(2), Sudjadi Sudjadi(3)

(1) Fakultas Pertanian Universitas Brawijaya
(2) Fakultas Farmasi Universitas Gadjah Mada
(3) Fakultas Farmasi Universitas Gadjah Mada
(*) Corresponding Author

Abstract


Bacillus thuringiensis is an entomopathogenic organism. The pathogenic effect is caused by crystalline protein, δ-endotoxin, which is encoded by the cry gene. The aim of this study was to clone the cry gene from B. thuringiensis var. kurstaki with a genomic library approach. HindIII digested B. thuringiensis total DNA was ligated to HindIII digested of pUC19, and then used to transform Escherichia coli DH5α. Selection of transformans carrying recombinant was done by α-complementation and a recombinant clone containing the cry gene was further screened by non-radioactive hybridization method using a probe synthesized from the conserved region of the published cry genes. The result suggested that two recombinant clones with the insert size of 3.4 kb and 2.0 kb, respectively, carrying the cry gene.


Keywords


cloning; cry gene; Bacillus thuringiensis

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References

Attathom, T., Chongrattanameteekul, W., Chanpaisang, I., & Siriyan, R. 1995. Morphological diversity and toxicity of delta-endotoxin produced by various strain of Bacillus thuringiensis, Bull. Entomo. Research. 85: 167-173.

Brown, T.A. 1986. Gene cloning. Van Nostrand Reinhold Co. Ltd., England; 234 pp.

Hofte, H. & Whiteley, . H.R. 1989. Insecticidal crystal proteins of Bacillus thuringiensis, Microbiol. Rev. 53: 242-255.

Kartono, G. 1991. Peranan Gossipol dalam ketahanan tanaman kapas terhadap Helicoverpa armigera (Hubner) Hardwick) (Lepidoptera Noctuidae), Disertasi dalam Ilmu Petanian, UGM. Yogyakarta: 160.

Llewellyn, D.J., Brown, M. Cousins, Y., Hartweck, L., Last. D., Mathews, A., Murray, F. & Thisleon, J. 1992. The science behind transgenic cotton plant, Proc. 1992. Australian Cotton Conference: 265-379.

Roy, P. 1992. Genomic amplification and expression of δ-endotoxin fragment of Bacillus thuringiensis, Biochem. Biophys. Research. Comm. 182: 641-647.

Sambrook, J., Fritsch,E.F., & Maniatis,T. 1989. Molecular Cloning. A Laboratory Manual, Second Edition, Cold Spring Harbor Laboratory Press.

Schnepf, H.E., Wong, H.C. & Whiteley, H.R. 1985. The amino acid sequence of a crystal protein from Bacillus thuringiensis deduced from the DNA base sequence, J.BioI. Chem. 260: 6264-6272.

Starkey, M.P., Fenning, T., Davey, M.R. & Mulligan, 1991. Desing and use of synthetic oligonucleotide probes in the cloning of d-endotoxin genes from Bacillus thuringiensis, Enzyme Microb. Technol. 13: 661-664.

Wong, H.C. & Whiteley, H.R. 1985. The amino acid sequence of a crystal protein from Bacillus thuringiensis deduced from the DNA base sequence, J.Biol.Chem., 260: 6264-6272.



DOI: https://doi.org/10.22146/jpti.9918

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