• Molecules and Cells
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• 제40권11호
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• pp.814-822
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• 2017

Meiotic homologous recombination generates new combinations of preexisting genetic variation and is a crucial process in plant breeding. Within the last decade, our understanding of plant meiotic recombination and genome diversity has advanced considerably. Innovation in DNA sequencing technology has led to the exploration of high-resolution genetic and epigenetic information in plant genomes, which has helped to accelerate plant breeding practices via high-throughput genotyping, and linkage and association mapping. In addition, great advances toward understanding the genetic and epigenetic control mechanisms of meiotic recombination have enabled the expansion of breeding programs and the unlocking of genetic diversity that can be used for crop improvement. This review highlights the recent literature on plant meiotic recombination and discusses the translation of this knowledge to the manipulation of meiotic recombination frequency and location with regards to crop plant breeding.

• 미생물학회지
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• 제30권2호
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• pp.83-87
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• 1992

선택적 표식인자가 전위인자일때, 표식인자의 선택 순서에 따라 동시형질도가 달라진다. 본 연구에서는 이 현상을 설명하기 위하여 아래와 같은 수학적 접근을 시도하였다. 먼저, transducing particle 들의 형성과정을 다섯종류로 유별하였다. 그런다음, 하나의 표지가 고정될 가능성을 의미하는 확률함수들을 확률밀도 개념에 근거하여 수학적으로 정식화였다. 그리고 실제 값들과 유용한 가정들을 이식들에 결합하여 그로부터 얻어진 이론적인 빈도를 실측 값들과 비교하였다. 이려한 분석에 의하여 그 빈도의 상이성이 일반 재조합에 필요한 상용성을 전위인자가 제공하지 봇하고, 그 주변에 재조합 빈도를 거리에 의존하는 형태로 감소시키기 때문이라는 결론을 얻었다.

• ETRI Journal
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• 제30권1호
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• pp.89-100
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• 2008

We consider the feature recombination technique in a multiband approach to speaker identification and verification. To overcome the ineffectiveness of conventional feature recombination in broadband noisy environments, we propose a new subband feature recombination which uses subband likelihoods and a subband reliable-feature selection technique with an adaptive noise model. In the decision step of speaker recognition, a few very low unreliable feature likelihood scores can cause a speaker recognition system to make an incorrect decision. To overcome this problem, reliable-feature selection adjusts the likelihood scores of an unreliable feature by comparison with those of an adaptive noise model, which is estimated by the maximum a posteriori adaptation technique using noise features directly obtained from noisy test speech. To evaluate the effectiveness of the proposed methods in noisy environments, we use the TIMIT database and the NTIMIT database, which is the corresponding telephone version of TIMIT database. The proposed subband feature recombination with subband reliable-feature selection achieves better performance than the conventional feature recombination system with reliable-feature selection.

• Journal of Microbiology and Biotechnology
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• 제18권3호
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• pp.585-590
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• 2008

Human tropic Porcine Endogenous Retroviruses (PERVs) are the major concern in zoonosis for xenotransplantation because PERVs cannot be eliminated by specific pathogen-free breeding. Recently, a PERV A/C recombinant with PERV-C bearing PERV-A gp70 showed a higher infectivity (approximately 500-fold) to human cells than PERV-A. Additionally, the chance of recombination between PERVs and HERVs is frequently stated as another risk of xenografting. Overcoming zoonotic barriers in xenotransplantation is more complicated by recombination. To achieve successful xenotransplantation, studies on the recombination in PERVs are important. Here, we cloned and sequenced proviral PERV env sequences from pig gDNAs to analyze natural recombination. The envelope is the most important element in retroviruses as a pivotal determinant of host tropisms. As a result, a total of 164 PERV envelope genes were cloned from pigs (four conventional pigs and two miniature pigs). Distribution analysis and recombination analysis of PERVs were performed. Among them, five A/B recombinant clones were identified. Based on our analysis, we determined the minimum natural recombination frequency among PERVs to be 3%. Although a functional recombinant envelope clone was not found, our data evidently show that the recombination event among PERVs may occur naturally in pigs with a rather high possibility.

• The Plant Pathology Journal
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• 제24권1호
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• pp.1-7
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• 2008

Fusarium verticillioides (teleomorph Gibberella moniliformis) is associated with maize worldwide causing ear rot and stalk rot, and produces fumonisins, a group of mycotoxins detrimental to humans and animals. While research tools are available, our understanding of the molecular mechanisms associated with fungal virulence and fumonisin biosynthesis in F. verticillioides is still limited. One of the restraints that hampers F. verticillioides gene characterization is the fact that homologous recombination (HR) frequency is very low (<2%). Screening for a true gene knock-out mutant is a laborious process due to a high number of ectopic integrations. In this study, we generated a F. verticillioides mutant (SF41) deleted for FvKU70, a gene directly responsible for non-homologous end-joining mechanism, with the aim of improving HR frequency. Here, we demonstrate that FvKU70 deletion does not impact key Fverticillioides phenotypes, e.g., development, secondary metabolism, and virulence, while dramatically improving HR frequency. Significantly, we also confirmed that a high percentage (>85%) of the HR mutant strains harbor a desired mutation with no additional copy of the mutant allele inserted in the genome. We conclude that SF41 is suitable for use as a type strain when performing high-throughput gene function studies in F. verticillioides.

• Molecules and Cells
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• 제45권5호
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• pp.273-283
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• 2022

During meiosis, homologous chromosomes (homologs) pair and undergo genetic recombination via assembly and disassembly of the synaptonemal complex. Meiotic recombination is initiated by excess formation of DNA double-strand breaks (DSBs), among which a subset are repaired by reciprocal genetic exchange, called crossovers (COs). COs generate genetic variations across generations, profoundly affecting genetic diversity and breeding. At least one CO between homologs is essential for the first meiotic chromosome segregation, but generally only one and fewer than three inter-homolog COs occur in plants. CO frequency and distribution are biased along chromosomes, suppressed in centromeres, and controlled by pro-CO, anti-CO, and epigenetic factors. Accurate and high-throughput detection of COs is important for our understanding of CO formation and chromosome behavior. Here, we review advanced approaches that enable precise measurement of the location, frequency, and genomic landscapes of COs in plants, with a focus on Arabidopsis thaliana.

• 미생물학회지
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• 제41권4호
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• pp.239-245
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• 2005

Transformation-associated recombination (TAR) 클로닝 법은 복잡한 게놈으로부터 염색체 내의 특정부위나 유전자를 선택적으로 분리할 수 있다. 이 방법은 목적 유전자에 근접한 작은 게놈DNA 염기서열 정보를 필요로 한다. 이 기술은 효모의 spheroplast transformation을 시키는 동안 목적으로 하는 유전자의 5' 또는 3' 서열을 포함하고 있는 TAR vector와 게놈DNA사이에서 일어나는 상동성재조합에 의해 이루어진다. 본 연구에서는 plasmid 모델시스템을 이용하여 target hooks 내에 존재하는 비상동성 염기서 열이 상동성재조합에 미치는 영향을 조사하였다. plasmid에 존재하는HIS3유전자와 변형시킨 his3-TRP1-his3 단편 사이의 상동성재조합의 효율은 $Ura^+$ 형질전환체의 형질분석에 의해 이루어졌다. $Ura^+$ 형질전환체의 수는 7종류의 서로 달리 변형된 his3-TRP1-his3 단편들을 사용하였을 매 거의 동일하게 나타났다. 그러나 $Trp^+His^+$ positive recombinants의 빈도는 변형된 his3-TRP1-his3 단편 내에 비상동성 영역에 부정확한 간격을 지닐 때 현저한 감소를 나타내었다. 이러한 결과로서, 부정확한 간격이 target hook과 substrate DNA 사이에 일어나는 상동성재조합을 방해하는 것으로 사료된다. 그러므로 이종간의 상동유전자를 클로닝 할 때에는 target hook내의 비상동성 염기서열이 존재한다면 이것이 정확한 간격을 지니는지 여부를 중요란 요인으로 고려해야 한다.

• 한국미생물·생명공학회지
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• 제14권3호
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• pp.265-270
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• 1986

L. casei세포의 유전연구를 위한 도구로서 세포융합 기술을 연구하였으며 융합세포(recombinant)를 선발하고 확인하기 위한 유전자 선발표지 인자로서 항생제 저항성이 이용되었고, 항생제 저항성 돌연변이 균주는 nitrosoguanidine을 처리하여 분리하였다. 선발 배지에서 항생제의 적절한 최종 농도는 streptomycin 25 $\mu\textrm{g}$/$m{\ell}$, hostacillin 0.5 I. U./$m{\ell}$, lincomycin 0.5$\mu\textrm{g}$/$m{\ell}$ 그리고 methicillin 5 $\mu\textrm{g}$/$m{\ell}$로 확인되었다. L. casei균주에서 높은 세포융합은 PEG 분자량 4,000에서 40%농도, 중성 부근의 pH, 3$0^{\circ}C$에서 약 1분간 처리하였을 때 얻어 졌다. 항생제 저항성의 자연돌연변이주의 출현빈도는 세포융합 출현빈도 보다 $10^2$-$10^3$ 정도 낮은 수준으로 나타났다. 세포융합 빈도는 모균에 대해 약 $10^{-4}$ 비율로 나타났다.

• Journal of Microbiology and Biotechnology
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• 제27권2호
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• pp.335-341
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• 2017

The complexity of the bacterial recombination system is a barrier for the construction of bacterial mutants for the further functional investigation of specific genes. Several protocols have been developed to inactivate genes from the genus Pseudomonas. Those protocols are complicated and time-consuming and mostly do not enable easy construction of multiple knock-ins/outs. The current study describes a single and double crossover-recombination system using an optimized vector-free allele-exchange protocol for gene disruption and gene replacement in a single species of the family Pseudomonadaceae. The protocol is based on self-ligation (circularization) for the DNA cassette which has been obtained by overlapping polymerase chain reaction (Fusion-PCR), and carries an antibiotic resistance cassette flanked by homologous internal regions of the target locus. To establish the reproducibility of the approach, three different chromosomal genes (ncRNA31, rpoN, rpoS) were knocked-out from the root-associative bacterium Pseudomonas stutzeri A1501. The results showed that the P. stutzeri A1501 mutants, which are free of any plasmid backbone, could be obtained via a single or double crossover recombination. In order to optimize this protocol, three key factors that were found to have great effect on the efficiency of the homologous recombination were further investigated. Moreover, the modified protocol does not require further cloning steps, and it enables the construction of multiple gene knock-in/out mutants sequentially. This work provides a simple and rapid mutagenesis strategy for genome editing in P. stutzeri, which may also be applicable for other gram-negative bacteria.

• Asian Pacific Journal of Cancer Prevention
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• 제14권12호
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• pp.7091-7094
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• 2013

DNA repair capacity is crucial in maintaining cellular functions and homeostasis. However, it can be altered based on DNA sequence variations in DNA repair genes and this may lead to the development of many diseases including malignancies. Identification of genetic polymorphisms responsible for reduced DNA repair capacity is necessary for better prevention. Homologous recombination (HR), a major double strand break repair pathway, plays a critical role in maintaining the genome stability. The present study was performed to determine the frequency of the HR gene XRCC3 Exon 7 (C18067T, rs861539) polymorphisms in Saudi Arabian population in comparison with epidemiological studies by "MEDLINE" search to equate with global populations. The variant allelic (T) frequency of XRCC3 (C>T) was found to be 39%. Our results suggest that frequency of XRCC3 (C>T) DNA repair gene exhibits distinctive patterns compared with the Saudi Arabian population and this might be attributed to ethnic variation. The present findings may help in high-risk screening of humans exposed to environmental carcinogens and cancer predisposition in different ethnic groups.