• Title/Summary/Keyword: Cry-B

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Bacillus thuringiensis as a Specific, Safe, and Effective Tool for Insect Pest Control

  • Roh, Jong-Yul;Choi, Jae-Young;Li, Ming-Sung;Jin, Byung-Rae;Je, Yeon-Ho
    • Journal of Microbiology and Biotechnology
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    • v.17 no.4
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    • pp.547-559
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    • 2007
  • Bacillus thuringiensis (Bt) was first described by Berliner [10] when he isolated a Bacillus species from the Mediterranean flour moth, Anagasta kuehniella, and named it after the province Thuringia in Germany where the infected moth was found. Although this was the first description under the name B. thuringiensis, it was not the first isolation. In 1901, a Japanese biologist, Ishiwata Shigetane, discovered a previously undescribed bacterium as the causative agent of a disease afflicting silkworms. Bt was originally considered a risk for silkworm rearing but it has become the heart of microbial insect control. The earliest commercial production began in France in 1938, under the name Sporeine [72]. A resurgence of interest in Bt has been attributed to Edward Steinhaus [105], who obtained a culture in 1942 and attracted attention to the potential of Bt through his subsequent studies. In 1956, T. Angus [3] demonstrated that the crystalline protein inclusions formed in the course of sporulation were responsible for the insecticidal action of Bt. By the early 1980's, Gonzalez et al. [48] revealed that the genes coding for crystal proteins were localized on transmissible plasmids, using a plasmid curing technique, and Schnepf and Whiteley [103] first cloned and characterized the genes coding for crystal proteins that had toxicity to larvae of the tobacco hornworm, from plasmid DNA of Bt subsp. kurstaki HD-1. This first cloning was followed quickly by the cloning of many other cry genes and eventually led to the development of Bt transgenic plants. In the 1980s, several scientists successively demonstrated that plants can be genetically engineered, and finally, Bt cotton reached the market in 1996 [104].

Effectiveness of various nonpharmacological analgesic methods in newborns

  • Kumar, Pancham;Sharma, Rakesh;Rathour, Sukhdev;Karol, Sunidhi;Karol, Mohit
    • Clinical and Experimental Pediatrics
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    • v.63 no.1
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    • pp.25-29
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    • 2020
  • Background: Pain during the developmental period may adversely affect developing neuronal pathways and result in adverse neurodevelopmental, cognitive, and behavioral effects in later life. Immunizations, e.g., hepatitis B vaccine (HBV), administered at birth are painful experiences to which neonates are universally subjected. Purpose: Here we aimed to study and compare the effectiveness of various nonpharmacological pain management methods in newborns to enable the development of safe and effective analgesic methods for newborns. Methods: This prospective study was conducted at a tertiary care hospital in the Himalayan region. Three hundred term healthy neonates were divided into 6 groups of 50 each. Groups 1-5 were intervention groups, patients of which received a nonpharmacological intervention (breastfeeding, nonnutritive sucking, rocking, 25% sucrose, or distilled water) before the intramuscular HBV, while patients in group 6 received no intervention. The pain response in each group after the HBV injection was assessed and compared using cry duration and Douleur Aigue Nveau-ne (DAN) score, a behavioral acute pain rating scale for newborns. Results: Cry duration was decreased in all intervention groups, significantly so in the sucrose (19.90 seconds), breastfeeding (31.57 seconds), and nonnutritive sucking (36.93 seconds) groups compared with controls (52.86 seconds). DAN scores decreased significantly (P<0.05) at one or more points i.e. 30, 60, or 120 seconds in the breastfeeding and 25% sucrose intervention groups compared with controls. Conclusion: Oral sucrose and nonnutritive sucking are simple yet underutilized nonpharmacological interventions that effectively reduce pain in newborns.

EARLY EVENTS OCCURRING DURING LIGHT SIGNAL TRANSDUCTION IN PLANTS AND FUNGI

  • Hasunuma, Kohji;Ogura, Yasunobu;Yabe, Naoto
    • Journal of Photoscience
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    • v.5 no.2
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    • pp.73-81
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    • 1998
  • Light signals constitute major factors in regulating gene expression and morphogenesis in plants and fungi. Phytochrome A and B were well characterized red and far-red light receptors in plants. Red light signals increased the phosphorylation of 18 kDa protein, which was identified to be nucleoside diphosphate (NDP) kinase. The NDP kinase catalyzed autophosphorylation and had a protein kinase activity similar to MAP (mitogen activated protein) kinase. As candidates for blue light photoreceptors, cDNAs for CRY1 and CRY2 were isolated. The N-teminal regions of these proteins showed a high hornology to DNA photolyase. The 120 kDa protein first detected in Pisurn sativurn, which showed blue light induced phosphorylation was also detected in Arabidopsis thaliana. The 120 kDa protein was encoded by the nphl gene, which regulated positive phototropism of the plant. In Neurospora crassa, blue light irradiation of the membrane fraction prepared from roycelia stimulated the phosphorylation of the 15 kDa protein, which was also identifmd to be an NDP kinase. Recent progress in understanding early events in light signal transduction mainly in Pisum sativum Alaska, Arabidopsis thaliana and Neurospora crassa was summarized.

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Prevalence of Bacillus cereus Group in Rice and Distribution of Enterotoxin Genes

  • Jang, Ji-Hyun;Lee, No-A;Woo, Gun-Jo;Park, Jong-Hyun
    • Food Science and Biotechnology
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    • v.15 no.2
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    • pp.232-237
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    • 2006
  • Bacillus cereus group comprising B. cereus, B. thuringiensis, and B. mycoides was differentiated by polymerase chain reaction (PCR) and colony morphology. Prevalence of B. cereus group in rice and distribution of enterotoxin genes were determined as possible food poisoning agents. PCR using primers targeted for gyrB and cry genes could distinguish B. thuringiensis from B. cereus, and B. mycoides was differentiated by rhizoid morphological characteristics on nutrient agar. Among 136 rice and their processed products, prevalence of B. cereus group was 40%. B. cereus group consisted of 54 B. cereus, 11 B. thuringiensis, and 1 B. mycoides. Major isolates were B. cereus, with B. thuringiensis detected up to 10% among edible rice tested. Five enterotoxin genes, hbl, nhe, bceT, entFM, and cytK, were broadly distributed among B. cereus group, especially in B. cereus and B. thuringiensis. Prevalence of B. cereus group in rice and enterotoxin distribution suggest B. thuringiensis and B. cereus are toxigenic strain that should be controlled in rice and its products.

Characterization of Two Type Crystal Proteins Produced by Transformed Bacillus thuringiensis NT0423 (형질전환된 Bacillus thuringiensis NT0423 균주에 의해 생산된 두가지 형태의 내독소 단백질의 특성)

  • 박현우;김호산;유용만;김상현;서숙재;강석권
    • Microbiology and Biotechnology Letters
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    • v.21 no.5
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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.

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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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    • v.34 no.2
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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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Analysis of right border flanking sequence in transgenic chinese cabbage harboring integrated T-DNA (Agrobacterium을 이용하여 형질전환시킨 배추에서 T-DNA Right Border 인접염기서열 분석)

  • Ahn, Hong-Il;Shin, Kong-Sik;Woo, Hee-Jong;Lee, Ki-Jong;Kim, Hyo-Sung;Park, Yong-Hwan;Suh, Seok-Cheol;Cho, Hyun-Suk;Kweon, Soon-Jong
    • Journal of Plant Biotechnology
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    • v.38 no.1
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    • pp.15-21
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    • 2011
  • We developed 14 transgenic lines of Chinese cabbage (Brassica rapa) harboring the T-DNA border sequences and CryIAc1 transgene of the binary vector 416 using Agrobacterium tumefaciens-mediated DNA transfer. Six lines had single copy cryIAc1 gene and four of them contained no vector backbone DNA. Of the left border (LB) flanking sequences six nucleotides were deleted in transgenic lines 416-2 and 416-3, eleven nucleotides in line 416-9, and 65 nucleotides including the whole LB sequences in line 416-17, respectively. And we defined 499 bp of genomic DNA (gDNA) of transformed Chinese cabbage, and blast results showed 96% homology with Brassica oleracea sequences. PCR with specific primer for the right border (RB) franking sequence revealed 834 bp of PCR product sequence, and it was consisted of 3' end of cryIAc1, nosterminal region and 52 bp of Chinese cabbage genomic DNA near RB. RB sequences were not found and the 58 nucleotides including 21 bp of nos-terminator 3' end were deleted. Also, there were deletion of 10 bp of the known genomic sequences and insertion of 65 bp undefined genomic sequences of Chinese cabbage in the integration site. These results demonstrate that the integration of T-DNA can be accompanied by unusual deletions and insertions both in transgenic and genomic sequences.

Microstructures fo Top Seed Milt-Processed $YBa_{2}$$Cu_{3}$$O_{7-\delta}$ Superconductor (종자결정법으로 성장시킨 $YBa_{2}$$Cu_{3}$$O_{7-\delta}$ 고온초전도체의 미세구조)

  • 한영희;성태현;한상철;이준성;정상진
    • Proceedings of the Korea Institute of Applied Superconductivity and Cryogenics Conference
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    • 1999.02a
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    • pp.43-46
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    • 1999
  • The microstructure of Top Seed Milt-Processed $YBa_{2}$$Cu_{3}$$O_{7-\delta}$ single crystal was studied. It was presumed that the segregation of Y211 is due to the difference of growth rates between a, b axis and c axis in crystal direction. Corn kernel lide structure which was grown by the diffusion of Y211 was observed. At the near corner of the seed crystal, the diagonal line on Y123 cry crystal is formed by the corn kernel like structure.

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Crystal Growth and Solid Solution of Hexagonal Ferrites (육방정 페라이트의 고용성 및 단결정 육성 연구)

  • 강진기;박병규;정수진
    • Journal of the Korean Ceramic Society
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    • v.23 no.3
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    • pp.78-86
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    • 1986
  • single crystals of various hexagonal ferrites were grown by a flux technique. For the growing experiment platinum crucibles of size 40 cc and a horizontal siliconit tube furnace were used. Charges consisted of the flux of BaO(SrO)/$B_2O_3$ and the composition of crystals in the system of BaO $(SrO)-Fe_2O_3-ZnO$. The BaO(SrO)/$B_2O_3$ molar ratio of the flux were varied from 1 to 3. Crystals up to 12.5mm in diameter were grown by slow cooling of melts from a maximum temperature of 1, 30$0^{\circ}C$or 1, 350$0^{\circ}C$ to 95$0^{\circ}C$ or 1, 00$0^{\circ}C$ The grown crystals exhibited a tabular hexagonal habits with very well developed ba-sal planes and narrow pyramidal faces of {1011} {1012} and {0001}. For the identification of the grown crystals X-ray diffraction studies were carried out. The effects of va-riations in flux ratio flux percentage and cooling rate on the quality of the grown crystals were studied. Cry-stal habits hillocks etch steps and growth spirals were observed by optical microscope. Magnetic properties of single crystals were measured.

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A simple model for selection and rapid advancement of transgenic progeny in sorghum

  • Visarada, K.B.R.S.;Saikishore, N.;Kuriakose, S.V.;Rani, V. Shobha;Royer, M.;Rao, S.V.;Seetharama, N.
    • Plant Biotechnology Reports
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    • v.2 no.1
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    • pp.47-58
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    • 2008
  • To select agronomically useful transgenic plants, a large number of transgenic events are initially produced, gene transfer confirmed, and advanced to obtain homozygous lines for testing in field trials. Direct in planta assays for identifying the transgene carriers in the segregating progeny are based on the activity of selectable marker gene and are easy, simple and inexpensive. For this purpose, expression of bar gene as measured by tolerance to damage by glufosinate ammonium, the active ingredient in the herbicide BASTA, was investigated. Dose damage curves were generated by leaf paint tests with BASTA on four genotypes of sorghum. Transgenic plants were characterized in terms of sensitivity to the concentration of glufosinate ammonium. In transgenics, symptoms of BASTA swab tests at different growth stages and PCR analysis for cry1B were carried out and correlated. Germination tests could not be employed for large scale evaluation of transgenic progeny because of mortality of tolerant seedlings after transplantation to soil. Based on the above findings, a simple, inexpensive, time-saving, two-step scheme for effective evaluation of transgenics and their progeny containing bar gene as selection marker using BASTA swab tests is described.