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논문 기본 정보
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- 저자정보
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- 한국잠사학회 International Journal of Industrial Entomology International Journal of Industrial Entomology 제18권 제1호
- 발행연도
- 2009.1
- 수록면
- 18 - 27 (10page)
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초록· 키워드
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B. thuringiensis strain LY-99 belonging to subsp. alesti(H3a3c), was isolated from Chinese tobacco warehouseand showed significantly high toxicity to Plutella xylostella.
For the identification of the cry1-type genes fromB. thuringiensis LY-99, an extended multiplex PCRrestrictionfragment length polymorphism (PCRRFLP)method was established by using two pairs ofuniversal primers based on the conserved regions ofthe cry1-type genes to amplify around 2.4 kb cry1-typegene fragments. Then the DNA fragment was clonedinto pGEM-T Easy vector and digested with EcoRIand EcoRV enzymes. Through this method, a knowncry1-type gene was successfully identified from the referencestrain, B. thuringiensis subsp. alesti. In addition,the RFLP patterns revealed that B. thuringiensisLY-99 included a novel cry1A-type gene in addition tocry1Aa, cry1Ac, cry1Be and cry1Ea genes. The novelcry1A-type gene was designated cry1Ah2 (Genbankaccession No DQ269474). An inverse PCR method wasused to amplify the flank regions of cry1Ah2 gene.
Finally, 3143 bp HindIII fragment from B. thuringiensisLY-99 plasmid DNA including 5’ region andpartial ORF was amplified, and sequence analysisrevealed that cry1Ah2 gene from LY-99 showed89.31% of maximum sequence similarity with cry1Ac1crystal protein gene. In addition, the deduced aminoacid sequence of Cry1Ah2 protein shared 87.80% ofmaximum identity with that of Cry1Ac2. This proteintherefore belongs to a new class of B. thuringiensiscrystal proteins.
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참고문헌 (22)
참고문헌 신청
Ben-Dov E. / 1997 / Extended screening by PCR for seven cry-group genes from field-collected strains of Bacillus thuringiensis / Appl Environ Microbiol 63 : 4883 ~ 4890
Ben-Dov E. / 1999 / Multiplex PCR screening to detect cry9 in Bacillus thuringiensis strains / Appl Environ Microbiol. 65 : 3714 ~ 3716
Ben-Dov E. / 2001 / PCR analysis of cry7 genes in Bacillus thuringiensis by the five conserved blocks of toxins / Curr Microbiol. 42 : 96 ~ 99
Birnboim JC. / 1979 / A rapid alkaline extraction procedure for screening recombinant plasmid DNA / Nucleic Acids Res. 7 : 1513 ~ 1523
Bourque SN. / 1993 / Multiplex polymerase chain reaction for detection and differentiation of the microbial insecticide Bacillus thuringiensis / Appl Environ Microbiol 59 : 5
Ben-Dov E. / 1999 / Multiplex PCR screening to detect cry9 in Bacillus thuringiensis strains / Appl Environ Microbiol. 65 : 3714 ~ 3716
Ben-Dov E. / 2001 / PCR analysis of cry7 genes in Bacillus thuringiensis by the five conserved blocks of toxins / Curr Microbiol. 42 : 96 ~ 99
Birnboim JC. / 1979 / A rapid alkaline extraction procedure for screening recombinant plasmid DNA / Nucleic Acids Res. 7 : 1513 ~ 1523
Bourque SN. / 1993 / Multiplex polymerase chain reaction for detection and differentiation of the microbial insecticide Bacillus thuringiensis / Appl Environ Microbiol 59 : 5
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