• Title/Summary/Keyword: OP-S3-1 primer

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Development of a psychrophilic-SCAR marker for Pleurotus eryngii (큰느타리버섯의 저온적응성 형질에 관련된 SCAR Marker 개발)

  • Kim, Su Chul;Hwang, Hye Sung;Cho, Yun Jun;Kim, Hye Su;Ryu, Jae-San;Cho, Soo Jeong
    • Journal of Mushroom
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    • v.11 no.3
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    • pp.171-176
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    • 2013
  • Genomic DNAs of psychrophilic strains of Pleurotus eryngii were analyzed by randomly amplified polymorphic DNA (RAPD) using OP-A, OP-B, OP-L, OP-P, OP-R and OP-S3 primers to develop the strain-specific DNA marker. A unique DNA fragment with the size of 480 bp was yielded by OP-S3 primer from the psychrophilic strain. A sequence characterized amplified region (SCAR) marker, designated as OP-S3-1, was designed on the basis of the determined sequence. The PCR analysis with the OP-S3-1 primer showed that this SCAR marker can clearly distinguish the psychrophilic strains from the control strains.

Development of strain-specific SCAR marker for selection of Pleurotus eryngii strains adaptable to high-temperature (큰느타리버섯의 고온적응성 형질에 관련된 SCAR Marker 개발)

  • Kim, Su Cheol;Kim, Hye Soo;Park, So Yeon;Ryu, Jae-San;Cho, Soo Jeong
    • Journal of Mushroom
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    • v.12 no.3
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    • pp.226-231
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    • 2014
  • In this study, SCAR marker that differentiates Pleurotus eryngii strains adaptable to high-temperature from control strain was developed. Genomic DNAs of 7 control strains of Pleurotus eryngii and 7 Pleurotus eryngii strains adaptable to high-temperature were analyzed by bulked segregant analysis (BSA) using randomly amplified polymorphic DNA (RAPD). Onehundred twenty RAPD primers were screened on bulked DNA samples and a unique DNA fragment with the size of 385 bp was yielded by OP-A06 primer from the Pleurotus eryngii strains adaptable to high-temperature. A sequence characterized amplified region (SCAR) marker, designated as OP-A06-1-F and OP-A06-1-R, was designed on the basis of the determined sequence. The PCR analysis with the OP-A06-1 primer showed that this SCAR marker can clearly distinguish the Pleurotus eryngii strains adaptable to high-temperature from the control strains.

Randomly Amplified Polymorphic DNA Analysis of Listeria Species Isolated from Foods in Korea (국내 식품으로부터 분리한 Listeria Species의 RAPD 분석)

  • 최영춘;박부길;이택수;오덕환
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.29 no.4
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    • pp.606-614
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    • 2000
  • This study was carried out for comparing Listeria strains developing genetic markers for Listeroa strains using Listeria sp. genetic markers using Randomly Amlymorphic DNA (RAPD) analysis method. Five of RAPD promers (OPA-01, OP-26-01, OP-26-02, OPB-01, OP-26-10) showed the distinctive polymorphism among Kisteria sp. isolated from domestic foods. RAPD-PCR with five arbitrary primers produced 76 DNA polymorphism. Among them, OPA-01 and OP-26-01 primers produced about 1.5kb and 0.7 kb amplified DNA fragments for all the Listeric relationships of Listeria sp. using NTSYS program were grouped into 7 clusters and showed 0.54 to 0.93 similarity among strains. Especially, No. 3 and No. 20 isolates showed the genetically most similar relationship by 0.94, and No. 7 and No. 24, or No. 7 and N0. 45 isolates showed the least similarty by 0.54 From these results, RAPD analysis method deemed to be successfully applied the classification and genetic analysis for Listeria sp. isolates.

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Cloning and Site-Directed Mutagenesis of Musca domestica Acetylcholinesterase for Enhancing Sensitivity to Organophosphorus and Carbamate Insecticides

  • Kim, Chung-Sei;Kim, Su-Il
    • Journal of Microbiology and Biotechnology
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    • v.16 no.11
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    • pp.1760-1772
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    • 2006
  • Mature acetylcholinesterase (AChE) gene (gm, 1,836 bp) was cloned from the housefly and successfully expressed in the E. coli CodonPlus (DE3) RIL system (GM-E, 72 kDa) with a yield of 1,630 mU/g fresh cells. Using the gm, 10 kinds of mutants were constructed and expressed for enhancing sensitivity to insecticides. The sensitivity of these mutants to five kinds of organophosphate (OP) and three carbamate insecticides was investigated by measuring the apparent bimolecular inhibition constant ($k_i=k_2/K_d$). Surprisingly, the sensitivity of quadruple mutant IGFT was enhanced as much as 7-fold for acephate, 164-fold for demeton-S-methyl, 484-fold for dichlorvos, 523-fold for edifenphos, 30-fold for ethoprophos, 30-fold for benfuracarb, 404-fold for carbaryl, and 107-fold for furathiocarb, compared with that of GM-E, although the sensitivity of each single point mutant was slightly increased. These mutational studies indicated that (i) contradictory to Walsh et al. [39], the residue 327 is the important key residue for enhancing sensitivity as much as the residue 262, (ii) the residue 82 and additional residues of 234, 236, and 585 are also important, and (iii) sensitivity was cooperatively accelerated as the number of strategic mutations increased.