• 제목/요약/키워드: Molecular genetics

검색결과 1,403건 처리시간 0.053초

Redesign of an Interhelical Loop of the Bacillus thuringiensis Cry4B delta-endotoxin for Proteolytic Cleavage

  • Krittanai, Chartchai;Lungchukiet, Panida;Ruangwetdee, Sarinthip;Tuntitippawan, Tipparut;Panyim, Sakol;Katzenmeier, Gerd;Angsuthanasombat, Chanan
    • BMB Reports
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    • 제34권2호
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    • pp.150-155
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    • 2001
  • The mosquito-larvicidal Cry4B protein from Bacillus thuringiensis subsp. israelensds was expressed in Escherichia coli. Upon activation by trypsin, the 130-kDa protoxin was processed into the 65-kDa active toxin containing two polypeptide fragments of ca. 47 and ca. 20 kDa. These two polypeptides are products of internal cleavages on the exposed loop connecting helices 5 and 6 in the seven-helical bundle domain. PCR-based mutagenesis was employed to introduce an additional cleavage site into the loop connecting helices 3 and 4. A series of amino acid changes were introduced into the targeted loop, resulting in seven mutant protoxins. Upon digestion with trypsin, a group of mutants with arginine introduced into the loop (EPRNQ, EPNRNQ, EPRNP, ESRNP and SSRNP) produced polypeptide products similar to those of the wild type (EPNNQ). When the loop, SSRNP, was expanded by an insertion of either asparagine (NSSRNP) or valine (VSSRNP), an additional cleavage was detected with proteolytic products of 47,12 and 6 kDa. This cleavage was confirmed to be at the introduced arginine residue by N-terminal sequencing. The mosquito larvicidal assay against Aedes aegypti demonstrated a relatively unchanged toxicity for the mutants without cleavage and reduced toxicity for those with an additional cleavage.

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Overview of citrin deficiency and its incidence in Asian region

  • Kobayashi, Keiko;Iijima, Mikio;Ushikai, Miharu;Lu, Yao Bang;Sheng, Jian-Sheng;Tabata, Ayako;Ikeda, Sayaka;Li, Meng Xian;Saheki, Takeyori;Okano, Yoshiyuki;Hsiao, Kwang-Jen;Hwu, Wuh-Liang;Yang, Yanling;Lau, Yu-Lung;Tsui, Lap-Chee;Choeh, Kyuchul;Lee, Dong-Hwan
    • 대한유전성대사질환학회지
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    • 제6권1호
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    • pp.89-93
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    • 2006
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Genomic Differentiation Among Oyster Mushroom Cultivars Released in Korea by URP-PCR Fingerprinting

  • Kang, Hee-Wan;Park, Dong-Suk;Park, Young-Jin;You, Chang-Hyun;Lee, Byoung-Moo;Eun, Moo-Yong;Go, Seong-Joo
    • Mycobiology
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    • 제29권2호
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    • pp.85-89
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    • 2001
  • URP primers of 20 mer derived from repetitive sequence of rice were used to assess genetic variation of oyster mushroom consisting of 10 cultivars of Pleurotus ostreatus, two cultivars of P. florida and two cultivars of P. sajor-caju which were registered in Korea. URP2F and URP38F primers produced cultivar-specific PCR polymorphic bands in the Pleurotus species. UPGMA cluster analysis using the URP-PCR data showed that 14 Pleurotus cultivars are genetically clustered into large three groups. The URP-PCR data provided important information for more efficient breeding strategies of Pleurotus cultivars.

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Trigger Factor Interacts with DnaA Protein to Stimulate its Interaction with DnaA Box

  • Lee, Yong-Sun;Lee, June;Kim, Hak-Kyun;Kang, Sukhyun;Han, Joo-Seok;Kim, Jae-Bum;Hwang, Deog-Su
    • Animal cells and systems
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    • 제7권1호
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    • pp.81-87
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    • 2003
  • While screening proteins that interact with DnaA protein, the initiator protein for Escherichia coil chromosomal DNA replication, we found a 52-kD sized protein which bound to DnaA protein in a salt-dependent manner. This protein was identified as trigger factor, a ribosome-associated peptidyl-prolyl- cisltrans isomerase with chaperone activity. Trigger factor was overproduced and purified to near homogeneity, and its effect on the function of DnaA protein was examined, Enhanced binding of DnaA protein to DnaA box with no apparent supershift in the gel-shift experiments suggested that trigger factor, by virtue of its chaperone activity, exerts a change on DnaA protein thus increasing its binding affinity for DnaA box.