• Title/Summary/Keyword: Progeny analysis

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Genome scan linkage analysis identifies a major quantitative trait loci for fatty acid composition in longissimus dorsi muscle in an F2 intercross between Landrace and Korean native pigs

  • Park, Hee-Bok;Han, Sang-Hyun;Yoo, Chae-Kyoung;Lee, Jae-Bong;Kim, Ji-Hyang;Baek, Kwang-Soo;Son, Jun-Kyu;Shin, Sang-Min;Lim, Hyun-Tae;Cho, In-Cheol
    • Asian-Australasian Journal of Animal Sciences
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    • v.30 no.8
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    • pp.1061-1065
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    • 2017
  • Objective: This study was conducted to locate quantitative trait loci (QTL) influencing fatty acid (FA) composition in a large $F_2$ intercross between Landrace and Korean native pigs. Methods: Eighteen FA composition traits were measured in more than 960 $F_2$ progeny. All experimental animals were genotyped with 165 microsatellite markers located throughout the pig autosomes. Results: We detected 112 QTLs for the FA composition; Forty seven QTLs reached the genome-wide significant threshold. In particular, we identified a cluster of highly significant QTLs for FA composition on SSC12. QTL for polyunsaturated fatty acid on pig chromosome 12 (F-value = 97.2 under additive and dominance model, nominal p-value $3.6{\times}10^{-39}$) accounted for 16.9% of phenotypic variance. In addition, four more QTLs for C18:1, C18:2, C20:4, and monounsaturated fatty acids on the similar position explained more than 10% of phenotypic variance. Conclusion: Our findings of a major QTL for FA composition presented here could provide helpful information to locate causative variants to improve meat quality traits in pigs.

Technical Development for Large DNA Fragment Transformation in Plants

  • Park, Su-Ryun;Seo, Mi-Suk;Lee, Sang-Kug;Park, Jee-Young;Kim, Hye-Ran;Lee, Hyo-Yeon;Bang, Jae-Wook;Lim, Yong-Pyo
    • Journal of Plant Biotechnology
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    • v.2 no.2
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    • pp.89-96
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    • 2000
  • For large DNA fragment transformation in dicots and monocots, BIBAC2 vector system was applied to Arabidopsis thaliana and Oryza sativa L. cv. Jinmi as a model plant, respectively. For Arabidopsis, the Th1 gene in T23L3 BAC clone whose size is about 90 kb was used as the target gene source for transformation. Because T23L3 BAC clone was originally constructed in pBelloBAC11, the target gene was reconstructed into BIBAC2. As the results of reconstruction, 476 colonies were survived in selection medium containing 40 mg/L kanamycin. In colony hybridization analysis, 24 out of 476 colonies exhibited positive signals. In the pulsed-field gel electrophoresis analysis, 11 out of 24 positive clones exhibited the band at the location of 90 kb. In Southern hybridization, positive signal band at the location of 90 kb was observed in all 11 transformants. Using these verified clones, Agrobacterium-mediated transformation was applied to Arabidopsis thaliana th1-201 mutant for genetic complementation test. Twelve thousands T$_1$ seeds were harvested, and antibiotic selection test is being analyzed to verify whether these seeds were transformed. for rice, COR356 that contains 150 kb human genomic DNA in a BIBAC2 vector was used as the target gene. As the results of transformation, 151 out of 210 co-cultivated calli were survived in selection medium containing 5 mg/L hygromycin, and 45 out of 151 survived calli were regenerated into plants. Transformation efficiency was 21.6%. Progeny test using 71 seeds is being analyzed now. These results provide the potential that large DNA fragments can be transferred into both dicots and monocot by Agrobacterium-mediate d transformation system.

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Expression of a Bovine ${\beta}$-Casein/Human Lysozyme Fusion Gene in the Mammary Gland of Transgenic Mice

  • Lee, Woon-Kyu;Kim, Sun-Jung;Hong, Seung-Beom;Lee, Tae-Hoon;Han, Yong-Mahn;Yoo, Ook-Joon;Im, Kyung-Soon;Lee, Kyung-Kwang
    • BMB Reports
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    • v.31 no.4
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    • pp.413-417
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    • 1998
  • Transgenic mice containing a bovine ${\beta}-Casein/Human$ lysozyme fusion gene (pBZ) were generated in order to produce human lysozyme in their milk. The expression vector was a quadripartite fusion consisting of a 2 kb upstream DNA of the bovine ${\beta}-casein$ gene, human lysozyme gene, intron II of the rabbit ${\beta}-globin$ gene, and the polyadenylation/termination signals of SV40 DNA. Fertilized mouse zygotes were microinjected with pBZ, then transferred into the oviduct of foster mothers. Out of 20 mice born, 11 survived until postweaning and three were identified as positivetransgenic by Southern blot analysis (one male and two females). The founder mice were mated to BCFl mice to produce transgenic progeny. It was confirmed by RT-PCR and Northern blot analyses that the transgene was specifically expressed in the mammary gland of the founder mice. Furthermore, the artificial introns within the transgenic RNA was proven to be correctly spliced out as judged by RT-PCR analysis. These results indicated that transgenic mice generated in this study properly expressed the human lysozyme RNA in their mammary gland.

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QTL Analysis of Teat Number Traits in an F2 Intercross between Landrace And Korean Native Pigs

  • Park, Hee-Bok;Han, Sang-Hyun;Yoo, Chae-Kyoung;Lee, Jae-Bong;Cho, Sang-Rae;Cho, In-Cheol
    • Journal of Embryo Transfer
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    • v.31 no.4
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    • pp.313-318
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    • 2016
  • The aim of this study was to identify quantitative trait loci (QTLs) influencing teat number traits in an $F_2$ intercross between Landrace and Korean native pigs (KNP). Three teat number traits (left;right;and total) were measured in 1105 $F_2$ progeny. All experimental animals were genotyped with 173 informative microsatellite markers located throughout the pig genome. We detect that seven chromosomes harbored QTLs for teat number traits: genome regions on SSC1;3;7;8;10;11;and 13. Six of fourteen identified QTL reached genome-wide significance. In SSC7;we identified a major QTL affecting total teat number that accounted for 5.6 % of the phenotypic variance;which was the highest test statistic (F-ratio = 61.1 under the additive model;nominal $P=1.3{\times}10^{-14}$) observed in this study. In this region;QTL for left and right teat number were also detected with genome-wide significance. With exception of the QTL in SSC10;the allele from KNP in all 6 identified QTLs was associated with decreased phenotypic values. In conclusion;our study identified both previously reported and novel QTL affecting teat number traits. These results can play an important role in determining the genetic structure underlying the variation of teat number in pigs.

Expression of Human Lactoferrin Gene in Transgenic Rice (Oryza sativa L.)

  • Lee, Yong-Eok;Oh, Seong-Eun;Nishiguchi, Satoshi;Riu, Key-Zung;Song, In-Ja;Park, Shin-Young;Lee, Jin-Hyoung;Kim, Il-Gi;Suh, Suk-Chul;Rhim, Seong-Lyul;Lim, Pyung-Ok;Lee, Hyo-Yeon
    • Journal of Plant Biotechnology
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    • v.34 no.2
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    • pp.145-152
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    • 2007
  • Lactoferrin is an 80-kDa iron-binding glycoprotein known to exert many biological activities, such as facilitating iron absorption and having antimicrobial and anti-inflammatory effects. Rice can be a useful target for edible food plants to introduce human lactoferrin, because it has lower allergenicity and is likely to be safer than microorganisms or transgenic animals. A cDNA fragment encoding human lactoferrin (HLF) driven by the maize polyubiquitin promoter, along with herbicide resistance gene (bar) driven by CaMV 35S promoter, was introduced into rice (Oryza sativa L. cv. Dong Jin) using the Agrobacterium -mediated transformation system. Putative transformants were initially selected on the medium containing bialaphos. The stable integration of the bar and HLF genes into transgenic rice plants was further confirmed through polymerase chain reaction (PCR) and Southern blot analyses. The expression of the full length HLF protein from various tissues such as grains and young leaves of transgenic rice was verified by Western blot analysis. Analysis of progeny also demonstrated that introduced genes were stably inherited to the next generation at the Mendelian fashion.

Identification and Functional Analysis of Mating Type Loci in the Pleurotus eryngii

  • Ryu, Jae San;Kim, Min-Keun;Park, Bokyung;Ali, Asjad;Joung, Wan-Kyu
    • 한국균학회소식:학술대회논문집
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    • 2015.11a
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    • pp.35-35
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    • 2015
  • Pleurotus eryngii has recently become a major cultivated mushroom; it uses tetrapolar heterothallism as a part of its reproductive process. Sexual development progresses only when the A and B mating types are compatible. Such mating incompatibility occasionally limits the efficiency of breeding programs in which crossing within loci-shared strains or backcrossing strategies are employed. Therefore, understanding the mating system in edible mushroom fungi will help provide a short cut in the development of new strains. We isolated and identified pheromone and receptor genes in the B3 locus of P. eryngii and performed a functional analysis of the genes in the mating process by transformation. A genomic DNA library was constructed to map the entire mating-type locus. The B3 locus was found to contain four pheromone precursor genes and four receptor genes. Remarkably, receptor PESTE3.3.1 has just 34 amino acid residues in its C-terminal cytoplasmic region; therefore, it seems likely to be a receptor-like gene. Real-time quantitative RT-PCR (real-time qRT-PCR) revealed that most pheromone and receptor genes showed significantly higher expression in monokaryotic cells than dikaryotic cells. The pheromone genes PEphb3.1 and PEphb3.3 and the receptor gene PESTE3.3.1 were transformed into P5 (A3B4). The transformants were mated with a tester strain (A4B4), and the progeny showed clamp connections and a normal fruiting body, which indicates the proposed role of these genes in mating and fruiting processes. This result also confirms that PESTE3.3.1 is a receptor gene. In this study, we identified pheromone and receptor genes in the B3 locus of P. eryngii and found that some of those genes appear to play a role in the mating and fruiting processes. These results might help elucidate the mechanism of fruiting differentiation and improve breeding efficiency.

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PRK1, a Receptor-like Kinase from Petunia inflata, is Essential for Post-meiotic Development of Pollen and Embryo Sac

  • Pai, Hyun-Sook;Karunanandaa, Balasulojini;Gilroy, Simon;Kao, Teh-Hui
    • Proceedings of the Botanical Society of Korea Conference
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    • 1996.07a
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    • pp.48-60
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    • 1996
  • We previously identified and characterized a predominantly pollen-expressed gene of Petunia inflata that encodes a receptor-like kinase named PRK1. The extracellular domain of PRK1 contains leucine-rich repeats which have been implicated in protein-protein interactions, and the cytoplasmic domain was found to autophosphorylate on serine and tyrosine. To investigate the function PRK1 in pollen development, we transformed P. inflata plants with a construct containing the promoter of a predominantly pollen-expressed gene of tomato, LAT52, fused to an antisense PRK1 cDNA corresponding to part of the extracellular domain of PRK1, There transgenic plants were found to each produce approximately equal amounts of normal and aborted pollen. Analysis of the inheritance of the transgene inserts in two of the transgenic plants, ASRK-13 and ASRK-20, to their progeny revealed that certain transgene inserts cosegregated with the pollen abortion phenotype. Microscopic examination of the aborted pollen grains showed that their outer wall, the exine, was essentially normal, but that their cytoplasm contained only starch-like granules. Staining of the nuclei of the microspores at different stages of uninucleate stage. However, at subsequent stages half of the microspores completed mitosis and developed into normal binucleate pollen, but the other half initially remained uninucleate, then lost their nucleio. Analysis of the amounts of PRK1 mRNA and the antisense PRK1 transcript suggested that the pollen abortion phenotype most likely resulted from down-regulation of the PRK1 gene by the antisense PRK1 transgene. These results suggest that PRK1 plays an essential role in a signal transduction pathway that mediates post-meiotic development of microspores.

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Mating Systems and Inbreeding Pressure in Populations of Wild Lentil Tare, Vicia tetrasperm (Leguminosae) (얼치기완두(콩과) 집단의 교배계와 내교잡 압력)

  • Huh, Man-Kyu
    • Journal of Life Science
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    • v.17 no.11
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    • pp.1477-1481
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    • 2007
  • The mating systems of natural populations of Vicia tetrasperm in Korea were determined using allozyme analysis. The result suggests that V. tetrasperm is low rates of outcrossing or mix-mating outcrossing (self-fertilization, s < 0.5). At the population levels, the values of inbreeding coefficient of ten populations in Korea varied from 0.131 to 0.176, giving an average 0.154. For ten natural populations, multi-locus estimates of outcrossing (tm) was 0.333 across fifteen polymorphic loci, with individual population values ranging from 0.269 to 0.423. The differences between the tm and ts values were not close to zero (tm - ts > 0.154), indicating that biparental inbreeding was significant in the loci. The reason for relatively low outcrossing rates of some populations could be attributed to extensive consanguineous mating and isolation of flowering mature plants. Although heterozygote excess was observed in one natural population, most populations exhibited varying degrees of inbreeding and heterozygotes deficit. Thus, selection against homozygotes operated in the progeny populations throughout the life cycle.

Fungal pathogen protection in transgenic lettuce by expression of a apoptosis related Bcl-2 gene (Apoptosis 관련 Bcl-2유전자의 도입을 통한 곰팡이 저항성 형질전환 상추의 육성)

  • Seo, Kyung-Sun;Min, Byung-Whan
    • Journal of Plant Biotechnology
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    • v.38 no.3
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    • pp.209-214
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    • 2011
  • Transgenic lettuce plants were successfully obtained from hypocotyl explants inoculated with Agrobacterium tumefaciens, which harbored a binary vector plasmid with Bcl-2 gene, related to apoptosis. After culture and selection on MS medium a number of kanamycin-resistant plantlets were regenerated. Polymerase chain reaction, Southern blot analysis and Northern blot analysis were used to identify and characterize the transgenic plants with the integrated Bcl-2 gene. Over 100 transgenic plants have been established in soil and flowered in the greenhouse. T1 progeny of 100 transgenic lettuce inbred lines were inoculated with Sclerotinia sclerotiorum. Expression of the Bcl-2 peptide in transgenic lettuce plants provides high levels of field resistance against Sclerotinia sclerotiorum, causal agent of the agronomically important fungal disease of lettuce.

Vascular Endothelial Growth Factor May Be Involved in the Behavioral Changes of Progeny Rats after Exposure to Ceftriaxone Sodium during Pregnancy

  • Yang, Xin;Tang, Ting;Li, Mengchun;Chen, Jie;Li, Tingyu;Dai, Ying;Cheng, Qian
    • Journal of Microbiology and Biotechnology
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    • v.32 no.6
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    • pp.699-708
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    • 2022
  • Antibiotic exposure during pregnancy have an adversely effects on offspring behavior and development. However, its mechanism is still poorly understood. To uncover this, we added ceftriaxone sodium to the drinking water of rats during pregnancy and conducted three-chamber sociability test, open-field test, and Morris water maze test in 3- and 6-week-old offspring. The antibiotic group offspring showed lower sociability and spatial learning and memory than control. To determine the role of the gut microbiota and their metabolites in the changes in offspring behavior, fecal samples of 6-week-old offspring rats were sequenced. The composition of dominant gut microbial taxa differed between the control and antibiotic groups. KEGG pathway analysis showed that S24-7 exerted its effects through the metabolic pathways including mineral absorption, protein digestion and absorption, Valine, leucine, and isoleucine biosynthesis. Correlation analysis showed that S24-7 abundance was negatively correlated with the level of VEGF, and metabolites associated with S24-7-including 3-aminobutanoic acid, dacarbazine, L-leucine, 3-ketosphinganine, 1-methylnicotinamide, and N-acetyl-L-glutamate-were also significantly correlated with VEGF levels. The findings suggest that antibiotic exposure during pregnancy, specifically ceftriaxone sodium, will adversely affects the behavior of offspring rats due to the imbalance of gut microbiota, especially S24-7, via VEGF and various metabolic pathways.