• Title/Summary/Keyword: Chromosomal Map

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Map based cloning of resistance to bacterial leaf blight gene using QTL analysis in rice

  • Du, Xiao-Xuan;Kim, Kyung-Min
    • Proceedings of the Korean Society of Crop Science Conference
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    • 2017.06a
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    • pp.138-138
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    • 2017
  • Agriculture is the most primitive civilized Activities of mankind but also the propellant of civilization development. Because it is the most basic material goods source of mankind. Among these materials rice is one of the most important part of these, we call them the substance of survival. From the beginning of the agricultural activities to the present we have experienced three industrial revolutions and are experiencing the Fourth Industrial Revolution. With the development of science and technology makes the efficiency of agricultural production is higher and higher, but compared with the original we are facing the same problem: natural disasters; pests and diseases; now also face the depletion of resources, environmental degradation and other issues. Therefore, improve and cultivate new crop varieties to make it better resistance and more production for better develop modern agriculture. It's very helpful for human social development. And also it is the responsibility and task of modern molecular breeding. In this study, I used bacterial leaf blight to find a better resistance gene to improve the resistance of rice. Frist Cultivate k3 of bacterial leaf blight, than inoculation by leaf clipping method (Kauffman,1973) in CNDH and SNDH population at 40days after rice transplanting. Check the lesion length by inoculation plants at 14days after inoculation, and record data for QTL analysis program. Than I get 4 intervals in 3 different chromosomal regions. I found these defense genes in the 4 intervals. So I used NCBI Justbio, Rapdb, etc. to finding these genes in physical map, than design primer for map base cloning. At last these defense genes will be employed in further research for introduction of the gene to the parental plant and rice breeding for solving food crisis.

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QTL Mapping of Genes Related with Grain Chemical Properties Based on Molecular Map of Rice

  • Kang, Hyeon-Jung;Cho, Yong-Gu;Lee, Young-Tae;Kim, Young-Doo;Eun, Moo-Young;Shim, Jae-Uk
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.43 no.4
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    • pp.199-204
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    • 1998
  • This study was conducted to investigate the chromosomal locations and effects of quantitative trait loci (QTL) associated with chemical properties of rice (Oryza sativa L.). One hundred sixty four recombinant inbred lines (MGRILs) of $F_{11}$ were derived from the cross between Milyang 23, Tongil type, and Gihobyeo, japonica type. They were evaluated for 7 traits of chemical property in rice. Transgressive segregation was observed for all traits examined. Eight significant QTLs were detected (LOD$\geq$2.0) for five traits, including two QTLs for amylose content, two QTLs for potassium content, one QTL for ratio of magnesium to potassium, one QTL for fat content and two QTLs for ash content. Phenotypic variation explained by each QTL ranged from 7.2% to 14.4%. However, no significant QTL was detected for magnesium and protein contents. In amylose content and ash content M alleles originated from Milyang 23 were responsible for increasing these traits and J alleles originated from Gihobyeo also responsible for increasing these traits. Pleiotropic effects of single QTLs on different traits are observed.

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Nucleotide Sequence and Properties of Macrolide-Lincosamide-Streptogramin B Resistance Gene from Staphylococcus aureus DH1 (Staphylococcus aureus DH1에서 분리된 Macrolide-Lincosamide-Streptogramin B 계열 항생물질에 대한 저항성 인자의 특성과 염기서열)

  • 권동현;박승문;윤권상;변우현
    • Korean Journal of Microbiology
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    • v.28 no.1
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    • pp.27-34
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    • 1990
  • Two macrolide-lincosamide-streptogramin B (MLS) antibiotic resistance genes, one expressed inducibly and the other expressed constitutively were recognized from a single Staphylococcus aureus DH1 strain. The inducible MLS resistance gene was isolated and cloned from the R-plasmid pDE1(7.4kb) and the constitutive gene was from chromosomal DNA. Base sequence of the inducible MLS resistance gene (1.2kb) was determined and found as same that of pE194. The restriction map of the cloned constitutive MLS resistance gene was compared with that of the inducible gene. Two genes have same restriction map except leader region. In the constitutive gene there is no leader region which is doing major role in inducible expression.

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Molecular Cloning and Expression of Bacillus pasteurii Urease Gene in Escherichia coli (B. pasteurii Urease 유전인자의 E. coli의 복제와 발현)

  • Kim, Sang-Dal;John Spizizen
    • Microbiology and Biotechnology Letters
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    • v.13 no.3
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    • pp.297-302
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    • 1985
  • The 7.1 Mdal Xbaf fragment of Bacillus pasteurii ATCC 11859 containing gene for urease was inserted into the Xbal site of bifunctional plasmid pGR71, and its urease gene was cloned and expressed in E. coil RRI. But the cloned gene was not expressed in Bacillus subtilis BR151 in consequence of deletion of inserted DNA fragment. The recombinant plasmid thus formed was named pGU66. The restriction map of the plasmid pGU66 was determined, and the size of the plasmid was estimated to be 12.6 Mdal by double digestion of restriction enzymes of the plasmid. The urease of the cloned strain was accumulated in periplasmic space and very similiar to that of donor strains in their enzymatic properties.

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Identification of QTLs Affecting Physical Traits of Cooked Rice

  • Kang, Hyeon-Jung;Cho, Yong-Gu;Lee, Young-Tae;Kim, Young-Doo;Eun, Moo-Young;Shim, Jae-Uk
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.44 no.1
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    • pp.70-73
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    • 1999
  • This study was conducted to ascertain the chromosomal locations and effect of quantitative trait loci (QTL) associated with the physical traits of rice (Oryza sativa L.) eating quality. One hundred sixty four recombinant inbred lines (MGRILs) of F$_{11}$ were derived from the cross between Milyang 23 (Tongil type) and Gihobyeo (japonica type). They were evaluated for six physical traits of cooked rice. Transgressive segregation was observed for all examined traits. Significant QTL were detected (LOD$\geq$2.0) in three traits, including single QTL for adhesiveness, gumminess, and chewiness of cooked rice, respectively. Phenotypic variation explained by each QTL ranged from 6.3% to 14.6%. However, no significant QTL was detected for hardness, cohesiveness, and elasticity of cooked rice. Pleiotropic effects of single QTL on different traits are observed.d.

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Localization of 5,105 Hanwoo (Korean Cattle) BAC Clones on Bovine Chromosomes by the Analysis of BAC End Sequences (BESs) Involving 21,024 Clones

  • Choi, Jae Min;Chae, Sung-Hwa;Kang, Se Won;Choi, Dong-Sik;Lee, Yong Seok;Park, Hong-Seog;Yeo, Jung-Sou;Choi, Inho
    • Asian-Australasian Journal of Animal Sciences
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    • v.20 no.11
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    • pp.1636-1650
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    • 2007
  • As an initial step toward a better understanding of the genome structure of Korean cattle (Hanwoo breed) and initiation of the framework for genomic research in this bovine, the bacterial artificial chromosome (BAC) end sequencing of 21,024 clones was recently completed. Among these clones, BAC End Sequences (BESs) of 20,158 clones with high quality sequences (Phred score ${\geq}20$, average BES equaled 620 bp and totaled 23,585,814 bp), after editing sequencing results by eliminating vector sequences, were used initially to compare sequence homology with the known bovine chromosomal DNA sequence by using BLASTN analysis. Blast analysis of the BESs against the NCBI Genome database for Bos taurus (Build 2.1) indicated that the BESs from 13,201 clones matched bovine contig sequences with significant blast hits (E<$e^{-40}$), including 7,075 single-end hits and 6,126 paired-end hits. Finally, a total of 5,105 clones of the Korean cattle BAC clones with paired-end hits, including 4,053 clones from the primary analysis and 1,052 clones from the secondary analysis, were mapped to the bovine chromosome with very high accuracy.

The transposition pattern of the Ac element and its use for targeted transposition in Arabidopsis thaliana

  • Machida, Yasunori;Onouchi, Hitoshi;Tanaka, Hirokazu;Hamada, Susumu;Ishikawa, Takaaki;Semiarti, Endang;Iwakawa, Hidekazu;Nomura, Kiyohito;Machida, Chiyoko
    • Proceedings of the Botanical Society of Korea Conference
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    • 1999.07a
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    • pp.11-15
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    • 1999
  • In order to evlauate feasibility of the gene tagging by the maize transposable element Ac in heterologous plant systems, we have investigated physical distances and directions of transposition of the element in Arabidopsis thaliana and tobacco cultured cell line BY-2. We prepared a T-DNA construct that carried a non-autonomous derivative of Ac with a site for cleavage by endonuclease I-Scel (designated dAc-I-RS element). Another cleavage site was also introduced into the T-DNA region outside dAc-I-RS. A number of transgenic Arabidopsis plants were generated, each of which had a single copy of the T-DNA at a different chromosomal location. To examine the pattern of transposition, three out of these transgenic plants were crossed with the Arabidopsis plant that carried the gene for Ac transposase and progeny in which dAc-I-RS had been transposed were isolated. After digestion of the genomic DNA of these progeny with I-SceI, sizes of segment of DNA were determined byd pulse-field gel electrophoresis. We also performed linkage analysis for the transposed elements and sites of mutations near the elements. Our results with three transgenic lines showed that 50% of all transposition events had occurred within 1,700 kilo-base pairs (kb) on the same chromosome, with 35% within 200 kb, and that the elements transposed in both directions on the chromosome with roughly equal probability. The data thus indicate that the Ac-Ds system is most useful for tagging of genes that are present within 200 kb of the chromosomal site of Ac in Arabidopsis. In addition, determination of the precise localization of the transposed dAc-I-RS element should definitely assist in map-based cloning of genes around insertion sites. In the present paper, we report typical examples of such gene isolation studies.

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세포내의 물의 상태

  • 강사욱
    • Proceedings of the Botanical Society of Korea Conference
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    • 1985.08b
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    • pp.51-57
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    • 1985
  • In order to evlauate feasibility of the gene tagging by the maize transposable element Ac in heterologous plant systems, we have investigated physical distances and directions of transposition of the element in Arabidopsis thaliana and tobacco cultured cell line BY-2. We prepared a T-DNA construct that carried a non-autonomous derivative of Ac with a site for cleavage by endonuclease I-Scel (designated dAc-I-RS element). Another cleavage site was also introduced into the T-DNA region outside dAc-I-RS. A number of transgenic Arabidopsis plants were generated, each of which had a single copy of the T-DNA at a different chromosomal location. To examine the pattern of transposition, three out of these transgenic plants were crossed with the Arabidopsis plant that carried the gene for Ac transposase and progeny in which dAc-I-RS had been transposed were isolated. After digestion of the genomic DNA of these progeny with I-SceI, sizes of segment of DNA were determined byd pulse-field gel electrophoresis. We also performed linkage analysis for the transposed elements and sites of mutations near the elements. Our results with three transgenic lines showed that 50% of all transposition events had occurred within 1, 700 kilo-base pairs (kb) on the same chromosome, with 35% within 200 kb, and that the elements transposed in both directions on the chromosome with roughly equal probability. The data thus indicate that the Ac-Ds system is most useful for tagging of genes that are present within 200 kb of the chromosomal site of Ac in Arabidopsis. In addition, determination of the precise localization of the transposed dAc-I-RS element should definitely assist in map-based cloning of genes around insertion sites. In the present paper, we report typical examples of such gene isolation studies.

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Molecular Cloning of $\beta$-Galactosidase Gene from Neisseria lactamica 2118 into Escherichia coli MC 1061 (Neisseria lactamica 2118의 $\beta$-galactosidase 유전자의 대장균으로의 클로닝)

  • Lee, Jong-Su
    • The Journal of Natural Sciences
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    • v.5 no.1
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    • pp.37-45
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    • 1992
  • The gene coding for $\beta$-galactosidase of Neisseria lactamica 2118 was cloned into Escherichia coli MC 1061. The isolated 6.5 Kb EcoR I fragement and 7.2 Kb BamH I fragment of chromosomal DNA in Southern hybridization were ligated to a vector plasmid pBR322 and then transformed into Escherichia coli MC 1061 cells. Finally, I obtained three clones as $\beta$-galactosidase positive clone by colony hybridization and Southern hybridization($\beta$-galactosidase probe: lac Z gene of pMC1871). Three recombinant plasmids(pNL.13. 17 and 24) were found to contain the 7.2Kb BamH I fragment originated from Neisseria lactamica 2118 chromosomal DNA by Southern hybridization and pNL 24 was showed high homology to probe especially and also its physical map was constructed.

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Chromosomal Localization of Korean Cattle (Hanwoo) BAC Clones via BAC end Sequence Analysis

  • Chae, Sung-Hwa;Kim, Jae-Woo;Choi, Jae Min;Larkin, Denis M.;Everts-van der Wind, Annelie;Park, Hong-Seog;Yeo, Jung-Sou;Choi, Inho
    • Asian-Australasian Journal of Animal Sciences
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    • v.20 no.3
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    • pp.316-327
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    • 2007
  • In this study, a Korean native cattle strain (Hanwoo) evidencing high performance in terms of both meat quality and quantity was employed in the generation of 150,000 BAC clones with an average insert size of 140 kb, and corresponding to about a 6X coverage of bovine chromosomal DNA. The BAC clones were pooled in a mini-scale via three rounds of a pooling protocol, and the efficiency of this pooling protocol was evaluated by testing the accuracy of accessibility to the positive clones, via a PCR-based screening method. Two sets of primers designed from each of two known genes were tested, and each yielded 2 or 3 positive clones for each gene, thereby indicating that the BAC library pooling system was appropriate with regard to the accession of the target BAC clones. Analyses of $3.3{\times}10^6$ base pairs obtained from the 7,090 BAC end sequence (BES) showed that 34.88% of the DNA sequence harbored the repetition sequence. Analysis of the 7,090 BES to the $1^{st}$ and $2^{nd}$ generation radiation hybrid map of the cattle genome, using the COMPASS program designed for the construction of a cattle-human comparative mapping, resulted in the localization of a total of 1,374 clones proximal to 339 $1^{st}$ generation markers, and 1,721 clones proximal to 664 $2^{nd}$ generation markers. Collectively, the BAC library and pooling system of the BAC clones from the Korean cattle, coupled with the chromosome-localized BAC clones, will provide us with novel tools for the excavation of desired clones for genome mapping and sequencing, and will also furnish us with additional information regarding breed differences in cattle.