• Title/Summary/Keyword: Gene Screening

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Detection of the Ryanodine Receptor Gene Mutation Associated with Porcine Stress Syndrome from Pig Hair Roots by PCR-RFLP (PCR-RFLP 기법을 이용한 Porcine Stress Syndrome의 진단)

  • Hwang, Eui-Kyung;Kim, Yeon-Soo
    • Korean Journal of Veterinary Research
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    • v.42 no.1
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    • pp.65-71
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    • 2002

  • We have utilized the PCR-RFLP method to detect the ryanodine receptor(RYR1) gene mutation and to estimate the genotype frequencies of the RYR1 gene in commercial crossbred pig population. The exon region(659bp) including point mutation(C ${\rightarrow}$T; Arg ${\rightarrow}$Cys) in the porcine ryanodine receptor gene, which is a causal mutation for PSS, was amplified by PCR and digested with Cfo I restriction enzyme. The RYR1 gene was classified into three genotypes by agarose gel electrophoresis. The normal homozygous(NN) individuals showed two DNA fragments consisted of 493 and 166bp. The mutant homozygous(nn) individuals showed only one DNA fragment of 659bp. Also, all three fragments(659, 493 and 166bp) were showed in heterozygous(Nn) carrier animals. The proportions of normal, carrier and PSS pigs within crossbred population of pigs were 81%, 15% and 4%, respectively. According to the results of analysis of variance for the association of genotypes of RYR1 of pigs at 30kg, day age at 90kg and average daily gains, the RYR1 nn genotype was very higher than RYR1 NN genotype for day age at 30kg with 5% level of significant difference, but no significant difference for association of any other genotypes with day age at 90kg and average daily gain in crossbred pigs. Therefore, DNA diagnosis by using PCR-RFLP analysis for the PSS gene was useful for large-scale screening of commercial pigs in the swine industry.

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Development of a Screening System for Drugs Against Human Papillomavirus-Associated Cervical Cancer: Based On E7-Rb Binding

  • Cho, Young-Sik;Cho, Cheong-Weon;Kang, Jeong-Woo;Cho, Min-Chul;Lee, Kyung-Ae;Shim, Jung-Hyun;Kwon, Our-Han;Choe, Yong-Kyung;Park, Sue-Nie;Yoon, Do-Young
    • BMB Reports
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    • v.34 no.1
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    • pp.80-84
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    • 2001

  • The human papillomavirus E7 protein can form a specific complex with a retinoblastoma tumor suppressor gene product (p105-Rb) that results in the release of the E2F transcription factor, which is critical for the growth-deregulation and transforming properties of the viral E7 oncoprotein. In an attempt to apply interaction between the E7 oncoprotein and a target cellular protein Rb for an in vitro screening system for drugs against human papillomavirus infection, we primarily investigated the E7Rb binding through a pull down assay and enzyme-linked immunosorbent assay. The pull down assay showed that both glutathione S-transferase-tagged E7 and His-tagged E7 immobilized on resins specifically produced complexes with bacterially expressed Rb in a dose-dependent manner, as determined by immunoblot analyses. This result coincided with that of an enzyme-linked immunosorbent assay, which is a useful system for the mass screening of potential drugs. Taken together, this screening system (based on the interaction between E7 and Rb) can be a promising system in the development of drugs against cervical cancers caused by human papillomavirus infection.

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Screening of melon genotypes identifies gummy stem blight resistance associated with Gsb1 resistant loci

  • Hassan, Md Zahid;Robin, Arif Hasan Khan;Rahim, Md Abdur;Natarajan, Sathishkumar;Kim, Hoy-Taek;Park, Jong-In;Nou, Ill-Sup
    • Journal of Plant Biotechnology
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    • v.45 no.3
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    • pp.217-227
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    • 2018

  • Gummy stem blight (GSB) is one of the most destructive and economically important, soil borne diseases of melon caused by the ascomycete fungus, Didymella bryoniae throughout the world. In Korea, however, no GSB resistant genotype has been reported yet. The study aimed to identify GSB resistant melon germplasm. We screened a total of 60 genotypes including 16 lines and 44 melon cultivars collected from USA and Korea. Among the 16 melon lines, four lines including 'PI482399', 'PI140471', 'PI136170' and 'PI420145', and two Korean cultivars viz. 'Asia Papaya' and 'Supra' showed complete resistance. We were aware that both genotypic and environmental variations could influence the phenotypic screening of resistance and susceptibility. We therefore, further assessed all genotypes using 20 SSR markers. The SSR marker 'CMCT505' linked to Gsb1 in chromosome 1 perfectly grouped resistant and susceptible lines indicating that resistance is probably due to the presence of Gsb1 gene. Cloning and sequencing of resistant and susceptible Gsb1 amplicons showed that there were 32-bp deletions in resistant line and 39-bp deletions in resistant cultivar compared to susceptible one. Thus, the resistant melon lines and cultivars identified in this study could be recommended for the melon breeding program. Furthermore, the SSR marker 'CMCT505' which is tightly linked with Gsb1 could be used for molecular screening of melon germplasm.

  • https://doi.org/10.5010/JPB.2018.45.3.217 인용 PDF KSCI