• 한국미생물·생명공학회지
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• 제25권2호
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• pp.159-165
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• 1997

We collected 3,237 plant samples, mainly leaves of various trees, from many provinces in Korea and a total of 1,925 Bacillus thuringiensis isolates were obtained and characterized. The isolates were characterized in terms of crystal morphology, PAGE pattern of the toxin proteins, plasmids pattern, biochemical characteristics, and bioassay. The microscopic observation showed that 49.1% of the isolates have bipyramidal shape crystals, 7.1% of spherical shape crystals, 1.4% of rhomboidal shape crystals, and others have small or amorphous inclusions. The insecticidal activities of the spore-crystal mixtures of isolates were tested against Plutella xylostella, Bombyx mori, Culex pipiens, and Agelastica coerulea. Bioassay showed that 51.3% of the isolates were shown to be active; lepidopteran-specific (44.8%), dipteran-specific(4.9%) and coleopteran-specific (1.6%). The remainder(48.8%) did not show any activity against the insects we tested. Interestingly though, some of these non-active isolates were shown to have bipyramidal crystals. By serotyping 22 isolates of our collection, we found that there are various kind of subspecies such as aizawai, amagiens, canadensis, darmstadiensis, galleriae, finitimus, kurstaki, morrisoni and neoleonensis, and three isolates have been classified into a new serotype, H49, and one of them, the type strain, named subsp. muju. From this study it was found that phylloplane is a good source for the isolation of Bacillius thuringiensis, and Bacillus thuringiensis is distributed widely in Korea.

• 한국응용곤충학회지
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• 제32권3호
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• pp.300-306
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• 1993

Bacillus thuringiensis subsp. kurstaki HD-1으로부터 생산된 살충성 내독소 단백질을 coding하는 CryIIA 유전자를 클로닝하고 염기서열을 조사하였다. HD-1 균주의 12개 plasmid 중 225kb plasmid를 분리하여 CryIIA 유전자를 포함하는 5kb HindIII 절편을 hybridization하여 찾아냈다. 이 절편을 plasmid pUC19에 ligation하여 E. coli에 형질 전환하였다. 이 독소 유전자를 포함하는 4kb BamHI-HindIII 절편은 vector pT7-5에 ligation하여 pSKIIA라하였다. pSKIIA는 3개의 open reading frames(orf1, orf2, orf3)로 구성되어 있으며 염기서열은 3,952base로 되어 있었다. 이러한 3개의 orf 각각의 발현 여부를 확인하기 위하여 생물검정을 하였다. 그러나 orf1 또는 orf2에 의한 형질 전환체는 독성이 없는 것으로 나타났다. orf3를 포함하는 형질 전환체는 3종의 나비목 곤충(배추점나방, 담배거세미나방, 담배나방) 및 1종의 파리목 곤충(집모기) 유충에 대하여 독성을 나타내었다.