• Title/Summary/Keyword: Evolutionary molecular engineering

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Simultaneous enhancement of thermostability and catalytic activity of phospholipase $A_1$ by evolutionary molecular engineering

  • Song, Jae-Kwang;Rhee, Joon-Shick
    • Proceedings of the Korean Society for Applied Microbiology Conference
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    • 2000.04a
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    • pp.168-171
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    • 2000
  • The thermal stability and catalytic activity of phospholipase A$_1$ from Serratia sp. MK1 were improved by an evolutionary molecular engineering. Two thermostable mutants were isolated after sequential rounds of error-prone PCR to introduce random mutations and filter-based screening of the resultant mutant library, and identified as having six (mutant TA3) and seven (mutant TA13) amino acid substitutions, respectively. Different types of the substitutions were found in two mutants, resulting in the increase of nonploar residues (mutant TA3) or changes between side chains within polar or charged residues (mutant TA13). The wild-type and mutant enzymes were purified, and the effect of temperature on their stability and catalytic activity was investigated. The T$\sub$m/ values of TA3 and TA13 were increased by 7 and 11$^{\circ}C$, respectively. Thus, evolutionary molecular engineering was found to be an effective and efficient approach to increasing thermostability without compromising enzyme activity.

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Evolutionary and Functional Analysis of Korean Native Pig Using Single Nucleotide Polymorphisms

  • Lee, Jongin;Park, Nayoung;Lee, Daehwan;Kim, Jaebum
    • Molecules and Cells
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    • v.43 no.8
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    • pp.728-738
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    • 2020
  • Time and cost-effective production of next-generation sequencing data has enabled the performance of population-scale comparative and evolutionary studies for various species, which are essential for obtaining the comprehensive insight into molecular mechanisms underlying species- or breed-specific traits. In this study, the evolutionary and functional analysis of Korean native pig (KNP) was performed using single nucleotide polymorphism (SNP) data by comparative and population genomic approaches with six different mammalian species and five pig breeds. We examined the evolutionary history of KNP SNPs, and the specific genes of KNP based on the uniqueness of non-synonymous SNPs among the used species and pig breeds. We discovered the evolutionary trajectory of KNP SNPs within the used mammalian species as well as pig breeds. We also found olfaction-associated functions that have been characterized and diversified during evolution, and quantitative trait loci associated with the unique traits of KNP. Our study provides new insight into the evolution of KNP and serves as a good example for a better understanding of domestic animals in terms of evolution and domestication using the combined approaches of comparative and population genomics.

PCR-mediated Recombination of the Amplification Products of the Hibiscus tiliaceus Cytosolic Glyceraldehyde-3-phosphate Dehydrogenase Gene

  • Wu, Linghui;Tang, Tian;Zhou, Renchao;Shi, Suhua
    • BMB Reports
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    • v.40 no.2
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    • pp.172-179
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    • 2007
  • PCR-mediated recombination describes the process of in vitro chimera formation from related template sequences present in a single PCR amplification. The high levels of genetic redundancy in eukaryotic genomes should make recombination artifacts occur readily. However, few evolutionary biologists adequately consider this phenomenon when studying gene lineages. The cytosolic glyceraldehyde-3-phosphate dehydrogenase gene (GapC), which encodes a NADP-dependent nonphosphorylating glyceraldehyde-3-phosphate dehydrogenase in the cytosol, is a classical lowcopy nuclear gene marker and is commonly used in molecular evolutionary studies. Here, we report on the occurrence of PCR-mediated recombination in the GapC gene family of Hibiscus tiliaceus. The study suggests that recombinant areas appear to be correlated with DNA template secondary structures. Our observations highlight that recombination artifacts should be considered when studying specific and allelic phylogenies. The authors suggest that nested PCR be used to suppress PCRmediated recombination.

Evolutionary Signature of Information Transfer Complexity in Cellular Membrane Proteomes

  • Kim, Jong-Min;Kim, Byung-Gee;Oh, S.-June
    • Genomics & Informatics
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    • v.7 no.2
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    • pp.111-121
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    • 2009
  • Cell membrane proteins play crucial roles in the cell's molecular interaction with its environment and within itself. They consist of membrane-bound proteins and many types of transmembrane (TM) proteins such as receptors, transporters, channel proteins, and enzymes. Membrane proteomes of cellular organisms reveal some characteristics in their global topological distribution according to their evolutionary positions, and show their own information transfer complexity. Predicted transmembrane segments (TMSs) in membrane proteomes with HMMTOP showed near power-law distribution and frequency characteristics in 6-TMS and 7-TMS proteins in prokaryotes and eukaryotes, respectively. This reaffirms the important roles of membrane receptors in cellular communication and biological evolutionary history.

Computational analysis of SARS-CoV-2, SARS-CoV, and MERS-CoV genome using MEGA

  • Sohpal, Vipan Kumar
    • Genomics & Informatics
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    • v.18 no.3
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    • pp.30.1-30.7
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    • 2020
  • The novel coronavirus pandemic that has originated from China and spread throughout the world in three months. Genome of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) predecessor, severe acute respiratory syndrome coronavirus (SARS-CoV) and Middle East respiratory syndrome coronavirus (MERS-CoV) play an important role in understanding the concept of genetic variation. In this paper, the genomic data accessed from National Center for Biotechnology Information (NCBI) through Molecular Evolutionary Genetic Analysis (MEGA) for statistical analysis. Firstly, the Bayesian information criterion (BIC) and Akaike information criterion (AICc) are used to evaluate the best substitution pattern. Secondly, the maximum likelihood method used to estimate of transition/transversions (R) through Kimura-2, Tamura-3, Hasegawa-Kishino-Yano, and Tamura-Nei nucleotide substitutions model. Thirdly and finally nucleotide frequencies computed based on genomic data of NCBI. The results indicate that general times reversible model has the lowest BIC and AICc score 347,394 and 347,287, respectively. The transition/transversions bias for nucleotide substitutions models varies from 0.56 to 0.59 in MEGA output. The average nitrogenous bases frequency of U, C, A, and G are 31.74, 19.48, 28.04, and 20.74, respectively in percentages. Overall the genomic data analysis of SARS-CoV-2, SARS-CoV, and MERS-CoV highlights the close genetic relationship.

In vitro Constructive Approaches to the Origin of Coding Sequences

  • Shiba, Kiyotaka
    • BMB Reports
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    • v.31 no.3
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    • pp.209-220
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    • 1998
  • How did nature create the first set of genes at the beginning of life on Earth? One of the goals of molecular biology is to elucidate the fundamental rules governing how genes and, therefore, proteins were created. Through experiments carried out in the emerging field of "in vitro" or "benchtop" evolution studies, we are gaining new insights into the origins of genes and proteins as well as the origins of their functions (e.g., catalysis). In this review, I present an overview of recent experimental approaches to the question of the origin and evolution of genes. In addition, I will introduce a novel in vitro protein emergence system that was recently developed in my laboratory.

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Bridging Comparative Genomics and DNA Marker-aided Molecular Breeding

  • Choi, Hong-Kyu;Cook, Douglas R.
    • Korean Journal of Breeding Science
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    • v.43 no.2
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    • pp.103-114
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    • 2011
  • In recent years, genomic resources and information have accumulated at an ever increasing pace, in many plant species, through whole genome sequencing, large scale analysis of transcriptomes, DNA markers and functional studies of individual genes. Well-characterized species within key plant taxa, co-called "model systems", have played a pivotal role in nucleating the accumulation of genomic information and databases, thereby providing the basis for comparative genomic studies. In addition, recent advances to "Next Generation" sequencing technologies have propelled a new wave of genomics, enabling rapid, low cost analysis of numerous genomes, and the accumulation of genetic diversity data for large numbers of accessions within individual species. The resulting wealth of genomic information provides an opportunity to discern evolutionary processes that have impacted genome structure and the function of genes, using the tools of comparative analysis. Comparative genomics provides a platform to translate information from model species to crops, and to relate knowledge of genome function among crop species. Ultimately, the resulting knowledge will accelerate the development of more efficient breeding strategies through the identification of trait-associated orthologous genes and next generation functional gene-based markers.

Comparative Genomics Reveals the Core and Accessory Genomes of Streptomyces Species

  • Kim, Ji-Nu;Kim, Yeonbum;Jeong, Yujin;Roe, Jung-Hye;Kim, Byung-Gee;Cho, Byung-Kwan
    • Journal of Microbiology and Biotechnology
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    • v.25 no.10
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    • pp.1599-1605
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    • 2015
  • The development of rapid and efficient genome sequencing methods has enabled us to study the evolutionary background of bacterial genetic information. Here, we present comparative genomic analysis of 17 Streptomyces species, for which the genome has been completely sequenced, using the pan-genome approach. The analysis revealed that 34,592 ortholog clusters constituted the pan-genome of these Streptomyces species, including 2,018 in the core genome, 11,743 in the dispensable genome, and 20,831 in the unique genome. The core genome was converged to a smaller number of genes than reported previously, with 3,096 gene families. Functional enrichment analysis showed that genes involved in transcription were most abundant in the Streptomyces pan-genome. Finally, we investigated core genes for the sigma factors, mycothiol biosynthesis pathway, and secondary metabolism pathways; our data showed that many genes involved in stress response and morphological differentiation were commonly expressed in Streptomyces species. Elucidation of the core genome offers a basis for understanding the functional evolution of Streptomyces species and provides insights into target selection for the construction of industrial strains.