• 한국잠사곤충학회지
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• 제35권2호
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• pp.129-133
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• 1993

Bacillus thruingiensis var. kurstaki HD-1의 내독소 단백질 유전자의 발현기작을 규명하기 위하여 이 균으로부터 내독소 단백질 유전자가 존재하는 것으로 확인된 29Md와 44Md plasmid를 분리한 후, Sau3AI 제어효소로 부분절단하고, pBR322 BamHI site에 ligation하여, E. coli HB101 strain에 transformation시켜, 3,000여개의 Ampr/Ters한 colony를 얻어 면역학적 방법과 살충적 검정으로 통해 재조합 균주 KC1을 얻었다. PKC1 plasmid DNA는 vector DNA를 포함하여 약 12kb 정도의 크기를 가지며, B. t k HD-1 내독소 단백질과 이동도가 같거나 일치하는 132kd, 117kd의 KC1 specific band 2개를 얻었다. KC1 cell extract를 첨식한 결과 약 80% 치사율을 보였다.

• Toxicological Research
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• 제29권3호
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• pp.149-155
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• 2013

G protein-coupled receptors (GPCRs) are membrane receptors; approximately 40% of drugs on the market target GPCRs. A precise understanding of the activation mechanism of GPCRs would facilitate the development of more effective and less toxic drugs. Heterotrimeric G proteins are important molecular switches in GPCR-mediated signal transduction. An agonist-activated receptor interacts with specific sites on G proteins and promotes the release of GDP from the $G{\alpha}$ subunit. Because of the important biological role of the GPCR-G protein coupling, conformational changes in the G protein upon receptor coupling have been of great interest. One of the most important questions was the interface between the GPCR and G proteins and the structural mechanism of GPCR-induced G protein activation. A number of biochemical and biophysical studies have been performed since the late 80s to address these questions; there was a significant breakthrough in 2011 when the crystal structure of a GPCR-G protein complex was solved. This review discusses the structural aspects of GPCR-G protein coupling by comparing the results of previous biochemical and biophysical studies to the GPCR-G protein crystal structure.

• Journal of Microbiology and Biotechnology
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• 제14권3호
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• pp.509-514
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• 2004

Six new serovarieties of B. thuringiensis carrying specific H-antigen have minor differences in biochemical characteristics and morphological characteristics of crystals, which are commonly resistant against four antibiotics. The B. thuringiensis serovar. coreanensis is nontoxic to silkworm larvae, but it is moderately toxic against the Culex pipiens larvae. The B. thuringiensis serovar. konkukian and leesis are nontoxic against mosquitos larvae, but are toxic against silkworm larvae. The B. thuringiensis serovar. seoulensis, sooncheon, and yosoo are highly toxic to B. mori larvae and moderately toxic to C. pipiens larvae. The six serovarieties harbor different plasmid DNA patterns. A 102-kDa protein is a major crystal protein in the four serovarieties and a 86-kDa protein is in one serovariety.

• Journal of Microbiology and Biotechnology
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• 제1권1호
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• pp.37-44
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• 1991

A shuttle promoter-probe vector, pEB203, was derived from pBR322, pPL703 and pUB110. Using the vector, a useful DNA fragment, 319 bp EcoRI fragment, having strong promoter activity has been cloned from Bacillus subtills chromosomal DNA. Selection was based on chloramphenicol resistance which is dependent upon the introduction of DNA fragments allowing expression of a chloramphenicol acetyl transferase gene. The nucleotide sequence of the 319 bp fragment has been determined and the putative -35 and -10 region, ribosome binding site, and ATG initiation codon were observed. This promoter was named EB promoter and the resultant plasmid which can be used as an expression vector was named pEBP313. The crystal protein gene from B. thuringiensis subsp. kurstaki HD-73 was cloned downstream from the EB promoter without its own promoter. When the resultant plasmid, pBT313, was introduced into Escherichia coli and B. subtilis, efficient synthesis of crystal protein was observed in both cells, and the cp gene expression in B. subtilis begins early in the vegetative phase. The cell extracts from both clones were toxic to Hyphantria cunea larvae.

• 한국미생물·생명공학회지
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• 제21권5호
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• pp.428-434
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• 1993

Cloning and expression of two different crystal protein genes from transformed Bacillus thuringiensis were investigated. B. thuringiensis NT0423 is toxic to both Lepidopterous and Dipte-rous larvae. The pCG5 vector carrying crystal protein genes (mosquitocidal and hemolytic activity) of B. thurigiensis subsp. morrisoni PG-14 was transformed into B. thurigiensis NT0423. Transformant has expressed two type crystals of bipyramid from NT0423 and ovoid from pCG5 in one cell.

• 한국잠사곤충학회지
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• 제40권2호
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• pp.126-130
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• 1998

A noninsecticidal strain, Bacillus thuringiensis NTB-88, isolated from Korean soil, had a typical bipyramidal parasporal inclusion and its serotype is identical to B. thuringiensis subspmorrisoni (H8a8b). To elucidate differences between insecticidal and noninsecticidal strains, we compared strain NTB-88 to other toxic B. thuringiensis subsp. morrisoni strains (HD-12 and PG-14). Restriction endonucleases digested plasmid DNA patterns showed that strain NTB-88 was different from lepidopteran-toxic strain, HD-12, but it was similar to dipteran-toxic strain, PG-14. The gene type of strain NTB-88 was different from those of other insecticidal strains, Furthermore, the NH2-terminal amino acid sequence of crystal protein of strain NTB-88 had no relation to those of the previously known $\delta$-endotoxins in other toxic strains as well as HD-12 and PG-14 strains. Therefore, the noninsecticidal crystal protein in strain NTB-88 is novel and its property is different from insecticidal ones.

• 한국미생물·생명공학회지
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• 제16권5호
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• pp.389-392
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• 1988

Escheriachia coli pSL 2-1 clone은 Bacillus sphaericus 1593의 모기살충 유전자를 클로닝한 재조합 DNA이다. 이 클론이 생산하는 살충독소 단백질의 분자량을 SDS-polyacrylamide gel을 이용하여 측정했다. B. sphaericus 1593균이 생산하는 독소결정체를 분리하여 전기영동을 한 결과는 6개의 단백질밴드(43, 58, 64, 100, 113, 130Kd)가 형성되었으나, 독소결정체를 알칼리 pH로 용해하여 전기영동을 하면 2개의 단백질 밴드(43과 64Kd)만이 나타났다. 그러나 대장균 pSL2-1균이 생산하는 독소단백질을 Sephadex G-200으로 정제하여 모기유충에 살충력이 있는 단백질을 전기영동한 결과는 42Kd만이 나타났다. LC50은 2 $\mu\textrm{g}$/$m\ell$이었다. B. sphaericus와 pSL2-1 clone 생산하는 살충단백질은 42Kd 단백질인 것으로 생각된다.

• 한국잠사곤충학회지
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• 제35권2호
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• pp.120-128
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• 1993

B. t k HD-1 균주에 0.002% SDS와 0.5$\mu$g/ml EtBr을 처리한 결과, 9개의 cry- 변이균주를 분리하였으며 또한 B. t k HD-1균주와 B. cereus 569균주를 혼합배양하는 방법으로 mating 실험을 수행하여 B. t k HD-1으로부터 일부 plasmid가 전이된 11개의 cry+ B. cereus와 2개의 cry=B. cereus를 분리하고 plasmid수와 분자량을 조사하였다. B. t k HD-1의 경우 9개의 plasmid가 존재하였고 일부 plasmid가 curing된 B. t k HD-1변이균주의 경우 29Md plasmid나 44Md plasmid가 반드시 존재하였으나, cry- 변이균주에는 29Md 이상의 모든 plasmid가 소실되어 내독소 단백질 합성에 관여하는 유전자가 기억된 plamid를 결정하였다.

• 한국응용곤충학회:학술대회논문집
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• 한국응용곤충학회 2002년도 추계 합동 특강 및 학술발표대회
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• pp.155-155
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• 2002

• 한국잠사학회:학술대회논문집
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• 한국잠사학회 2002년도 Join Meetings of Korean Society of Sericultural Science and Japanse Society of Sericultural Science
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• pp.114-114
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• 2002

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