• Title/Summary/Keyword: Insertional Variation

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Characterization of Insertional Variation of Porcine Endogenous Retroviruses in Six Different Pig Breeds

  • Jung, W.Y.;Yu, S.L.;Seo, D.W.;Jung, K.C.;Cho, I.C.;Lim, H.T.;Jin, D.I.;Lee, Jun-Heon
    • Asian-Australasian Journal of Animal Sciences
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    • v.25 no.10
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    • pp.1357-1363
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    • 2012
  • Pigs may need to be exploited as xenotransplantation donors due to the shortage of human organs, tissues and cells. Porcine endogenous retroviruses (PERVs) are a significant obstacle to xenotransplantation because they can infect human cells in vitro and have the potential for transmission of unexpected pathogens to humans. In this research, 101 pigs, including four commercial breeds (23 Berkshire, 13 Duroc, 22 Landrace and 14 Yorkshire pigs), one native breed (19 Korean native pigs) and one miniature breed (10 NIH miniature pigs) were used to investigate insertional variations for 11 PERV loci (three PERV-A, six PERV-B and two PERV-C). Over 60% of the pigs harbored one PERV-A (907F8) integration and five PERV-B (B3-3G, B3-7G, 742H1, 1155D9 and 465D1) integrations. However, two PERV-A loci (A1-6C and 1347C1) and one PERV-B locus (B3-7F) were absent in Duroc pigs. Moreover, two PERV-C loci (C2-6C and C4-2G) only existed in Korean native pigs and NIH miniature pigs. The results suggest that PERV insertional variations differ among pig breeds as well as among individuals within a breed. Also, the results presented here can be used for the selection of animals that do not have specific PERV integration for xenotransplantation research.

Trait Variation and Molecular Characterization of Ds insertional rice lines (벼 Ds 삽입변이 계통의 특성변이 및 분자생물학적 특성)

  • Ahn, Byung-Ohg;Kang, Kyung-Ho;Eun, Moo-Yong;Jeon, Yong-Hee;Yun, Doh-Won;Ji, Hyeon-So;Park, Sung-Han;Nam, Min-Hee;Suh, Seok-Cheol;Lee, Myung-Chul
    • Korean Journal of Breeding Science
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    • v.40 no.1
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    • pp.39-47
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    • 2008
  • The 1,874 rice lines were selected from 3,000 Ds insertional mutant pool by Basta herbicide treatment and were surveyed for trait variation and molecular characteristics of genes knocked out by Ds insertion. Compared with "Donjin", an original japonica cultivar used for transformation, Ds insertion mutant pool showed large variation in major agronomic traits including tiller, panicle, and heading etc. Southern blot analysis demonstrated that these lines on the average had two Ds copies in Donjin genome, resulting in 38.4% of one copy, 32.5% of two copies, 16.7% of three copies, and 11.3% of over four copies. GUS analysis showed that 3.9% of lines (73/1,860) had tissue-specific expression in leaves, nodal parts, floral organs such as stigma and pollen, and roots. Data set obtained from agricultural trait variation and molecular characteristics for individual Ds insertional lines would provide researchers with more information for understanding the function of unknown rice genes controlling economically important traits.

Agrobacterium-mediated transformation using gill tissue of Flammulina velutipes (Agrobacterium을 이용한 팽이 버섯 주름조직의 형질전환)

  • Park, Soon-Young;Van Peer, Arend F.;Jang, Kab-Yeul;Shin, Pyung-Gyun;Park, Yun-Hung;Yoo, Young-Bok;Park, Ki-Moon;Kong, Won-Sik
    • The Korean Journal of Mycology
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    • v.38 no.1
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    • pp.48-53
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    • 2010
  • Agrobacterium-mediated transformation was conducted in order to generate DNA insertional mutants of Flammulina velutipes. Agrobacterium tumefaciens AGL-1 harboring pBGgHg was transformed into gill tissues of Flammulina velutipes strain KACC42777. The transformants resistant on hygromycine ($30\;{\mu}g/ml$) were confirmed by PCR. The targeted insertional sites were amplified by inverse PCR and sequenced. To find the phenotype variation of all generated transformants, bottle cultivation which followed by the standard cultivation protocol were conducted. Color variation was observed on the cultivated fruiting bodies. Furthermore, the transformant pool will be used as a good genetic resources for studying gene function.

Analysis of Mutant Chinese Cabbage Plants Using Gene Tagging System (Gene Tagging System을 이용한 돌연변이 배추의 분석)

  • Yu, Jae-Gyeong;Lee, Gi-Ho;Lim, Ki-Byung;Hwang, Yoon-Jung;Woo, Eun-Taek;Kim, Jung-Sun;Park, Beom-Seok;Lee, Youn-Hyung;Park, Young-Doo
    • Horticultural Science & Technology
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    • v.28 no.3
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    • pp.442-448
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    • 2010
  • The objectives of this study were to analyze mutant lines of Chinese cabbage ($Brassica$ $rapa$ ssp. $pekinensis$) using gene tagging system (plasmid rescue and inverse polymerase chain reaction) and to observe the phenotypic characteristics. Insertional mutants were derived by transferring DNA (T-DNA) of $Agrobacterium$ for functional genomics study in Chinese cabbage. The hypocotyls of Chinese cabbage 'Seoul' were used to obtain transgenic plants with $Agrobacterium$ $tumefaciens$ harboring pRCV2 vector. To tag T-DNA from the Chinese cabbage genomic DNA, plasmid rescue and inverse PCR were applied for multiple copies and single copy insertional mutants. These techniques were successfully conducted to Chinese cabbage plant with high efficiency, and as a result, T-DNA of pRCV2 vector showed distinct various integration patterns in the transgenic plant genome. The polyploidy level analysis showed the change in phenotypic characteristics of 13 mutant lines was not due to variation in somatic chromosome number. Compared with wild type, the $T_1$ progenies showed varied phenotypes, such as decreased stamen numbers, larger or smaller flowers, upright growth habit, hairless leaves, chlorosis symptoms, narrow leaves, and deeply serrated leaves. The polyploidy level analysis showed the change in phenotypic characteristics of 13 mutant lines was not due to variation in somatic chromosome number. To tag T-DNA from the Chinese cabbage genomic DNA, plasmid rescue and inverse PCR were applied for multiple copies and single copy insertional mutants. Mutants that showed distinct phenotypic difference compared to wild type with 1 copy of T-DNA by Southern blot analysis, and with 2n = 20 of chromosome number were selected. These selected mutant lines were sequenced flanking DNA, mapped genomic loci, and the genome information of the lines is being recorded in specially developed database.

Sonoanatomic Variation of Pes Anserine Bursa

  • Imani, Farnad;Rahimzadeh, Poupak;Gharehdag, Farid Abolhasan;Faiz, Seyed Hamid Reza
    • The Korean Journal of Pain
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    • v.26 no.3
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    • pp.249-254
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    • 2013
  • Background: The pes anserine bursa lies beneath the pes anserine tendon, which is the insertional tendon of the sartorius, gracilis, and semitendinosus muscles on the medial side of the tibia, but it can lie in different sites in the medial knee. Accurate diagnosis of the position of the bursa is critical for diagnostic and therapeutic goals. The aim of this study was to evaluate sonoanatomic variations of the pes anserine bursa in the medial knee. Methods: One hundred seventy asymptomatic volunteers were enrolled in this study. Using ultrasound imaging (transverse approach, 7-13 MHz linear array probe) the sonoanatomic position of the pes anserine bursa and its relation to the pes anserine tendon were evaluated. Additionally, we evaluated the sonoanatomic variation of the saphenous nerve. Results: The position of the pes anserine bursa was between the medial collateral ligament and the pes anserine tendons in 21.2%/18.8% (males/females) of subjects; between the pes anserine tendons and the tibia in 67.1%/64.7% (m/f); and among the pes anserine tendons in 8.2%/12.9% (m/f). No significant differences in the position of the bursa existed between males and females. The saphenous nerve was found within the pes anserine tendons in 77.6%/74.1% (m/f) of subjects, but outside the pes anserine tendons in 18.8%/15.3% (m/f). Visibility of sonoanatomic structures was not related to either gender or BMI. Conclusions: Ultrasound provides very accurate information about variations in the pes anserine bursa and the saphenous nerve. This suggests that our proposed ultrasound method can be a reliable guide to facilitate approaches to the medial knee for diagnostic and therapeutic objectives.

Molecular Cloning and Alternative Splicing of Growth Hormone Transcripts in Greenling, Hexagrammos otakii (쥐노래미 (Hexagrammos otakii) 성장호르몬 cDNA유전자의 염기서열 변이 및 발현 특성)

  • Nam Yoon Kwon;Kim Dong Soo
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.35 no.6
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    • pp.676-681
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    • 2002
  • Different types of transcripts encoding growth hormone (GH) were identified from cDNA libraries constructed with pituitaries of a marine fish species, greenling (Hexagrammos otakii). GH-homologous cDNA clones were isolated using the high-density filter hybridization and the expressed sequence tag techniques. Of 39 full-length positive cDNA clones, 31 clones ($79\%$) displayed an identical sequence, however, remaining 8 clones exhibited several polymorphisms in their sequences including (1) the length and sequence variability in the 5' upstream region, (2) insertional sequences in open reading frame, and (3) deletion and/or single nucleotide polymorphism in the untranslated 3' region. Based on RT-PCT and RNA dot blot analyses, these transcripts were proven to be expressed in a pituitary-specific manner.