scholarly journals Development and Evaluation of Functional Gene Arrays for Detection of Selected Genes in the Environment

2001 ◽  
Vol 67 (12) ◽  
pp. 5780-5790 ◽  
Author(s):  
Liyou Wu ◽  
Dorothea K. Thompson ◽  
Guangshan Li ◽  
Richard A. Hurt ◽  
James M. Tiedje ◽  
...  
Keyword(s):  
Signal Intensity ◽  
Genomic Dna ◽  
Gene Diversity ◽  
Gene Array ◽  
Sequence Divergence ◽  
Quantitative Relationship ◽  
Gene Families ◽  
Functional Gene ◽  
Sequence Identity ◽  
Pure Cultures

ABSTRACT To determine the potential of DNA array technology for assessing functional gene diversity and distribution, a prototype microarray was constructed with genes involved in nitrogen cycling: nitrite reductase (nirS and nirK) genes, ammonia mono-oxygenase (amoA) genes, and methane mono-oxygenase (pmoA) genes from pure cultures and those cloned from marine sediments. In experiments using glass slide microarrays, genes possessing less than 80 to 85% sequence identity were differentiated under hybridization conditions of high stringency (65°C). The detection limit fornirS genes was approximately 1 ng of pure genomic DNA and 25 ng of soil community DNA using our optimized protocol. A linear quantitative relationship (r 2 = 0.89 to 0.94) was observed between signal intensity and target DNA concentration over a range of 1 to 100 ng for genomic DNA (or genomic DNA equivalent) from both pure cultures and mixed communities. However, the quantitative capacity of microarrays for measuring the relative abundance of targeted genes in complex environmental samples is less clear due to divergent target sequences. Sequence divergence and probe length affected hybridization signal intensity within a certain range of sequence identity and size, respectively. This prototype functional gene array did reveal differences in the apparent distribution ofnir and amoA and pmoA gene families in sediment and soil samples. Our results indicate that glass-based microarray hybridization has potential as a tool for revealing functional gene composition in natural microbial communities; however, more work is needed to improve sensitivity and quantitation and to understand the associated issue of specificity.

Elevation trend in bacterial functional gene diversity decouples from taxonomic diversity

CATENA ◽  
2021 ◽  
Vol 199 ◽  
pp. 105099
Author(s):  
Dorsaf Kerfahi ◽  
Ke Dong ◽  
Ying Yang ◽  
Hyoki Kim ◽  
Koichi Takahashi ◽  
...  
Keyword(s):  
Gene Diversity ◽  
Taxonomic Diversity ◽  
Functional Gene ◽  
Functional Gene Diversity

scholarly journals Sequence identity in an early chorion multigene family is the result of localized gene conversion.

Genetics ◽  
1991 ◽  
Vol 128 (3) ◽  
pp. 595-606
Author(s):  
B L Hibner ◽  
W D Burke ◽  
T H Eickbush
Keyword(s):  
Nucleotide Sequence ◽  
Gene Conversion ◽  
Nucleotide Sequence Identity ◽  
Early Period ◽  
Gene Families ◽  
Multigene Families ◽  
Coding Regions ◽  
Sequence Identity ◽  
Isolation And Characterization ◽  
Sequence Exchange

Abstract The multigene families that encode the chorion (eggshell) of the silk moth, Bombyx mori, are closely linked on one chromosome. We report here the isolation and characterization of two segments, totaling 102 kb of genomic DNA, containing the genes expressed during the early period of choriogenesis. Most of these early genes can be divided into two multigene families, ErA and ErB, organized into five divergently transcribed ErA/ErB gene pairs. Nucleotide sequence identity in the major coding regions of the ErA genes was 96%, while nucleotide sequence identity for the ErB major coding regions was only 63%. Selection pressure on the encoded proteins cannot explain this difference in the level of sequence conservation between the ErA and ErB gene families, since when only fourfold redundant codon positions are considered, the divergence within the ErA genes is 8%, while the divergence within the ErB genes (corrected for multiple substitutions at the same site) is 110%. The high sequence identity of the ErA major exons can be explained by sequence exchange events similar to gene conversion localized to the major exon of the ErA genes. These gene conversions are correlated with the presence of clustered copies of the nucleotide sequence GGXGGX, encoding paired glycine residues. This sequence has previously been correlated with gradients of gene conversion that extend throughout the coding and noncoding regions of the High-cysteine (Hc) chorion genes of B. mori. We suggest that the difference in the extent of the conversion tracts in these gene families reflects a tendency for these recombination events to become localized over time to the protein encoding regions of the major exons.

Cloning of taxane 2α-O-benzoyltransferase (TBT) genomic DNA from Taxus

Biologia ◽  
2006 ◽  
Vol 61 (3) ◽  
Author(s):  
Dongli Zhao ◽  
Le Li ◽  
Qian Wang ◽  
Guoyin Kai ◽  
Xiang Wang ◽  
...  
Keyword(s):  
Genomic Dna ◽  
Cdna Sequence ◽  
Gene Diversity ◽  
Genomic Sequences ◽  
Taxus Yunnanensis ◽  
Sequence Encoding ◽  
First Time

AbstractBased on the cDNA sequence encoding taxane 2α-O-benzoyltransferase (TBT) from Taxus yunnanensis (Gen-Bank Accession No.: AY970522), genomic sequences of TBTs from T. yunnanensis (TyTBT) and T. cuspidata (TcuTBT) were cloned for the first time. They both contain only one intron. The finding that the introns of TyTBT and TcuTBT are more diverse than their exons implies that the gene diversity is more within introns than within exons, which may be important for keeping the functions of the genes.

scholarly journals Microbial Functional Gene Diversity with a Shift of Subsurface Redox Conditions duringIn SituUranium Reduction

2012 ◽  
Vol 78 (8) ◽  
pp. 2966-2972 ◽  
Author(s):  
Yuting Liang ◽  
Joy D. Van Nostrand ◽  
Lucie A. N′Guessan ◽  
Aaron D. Peacock ◽  
Ye Deng ◽  
...  
Keyword(s):  
Functional Diversity ◽  
Field Experiments ◽  
Gene Diversity ◽  
Functional Gene ◽  
Redox Conditions ◽  
Functional Genes ◽  
Microbial Functional Diversity ◽  
Content Type ◽  
Microbial Functional Gene

ABSTRACTTo better understand the microbial functional diversity changes with subsurface redox conditions duringin situuranium bioremediation, key functional genes were studied with GeoChip, a comprehensive functional gene microarray, in field experiments at a uranium mill tailings remedial action (UMTRA) site (Rifle, CO). The results indicated that functional microbial communities altered with a shift in the dominant metabolic process, as documented by hierarchical cluster and ordination analyses of all detected functional genes. The abundance ofdsrABgenes (dissimilatory sulfite reductase genes) and methane generation-relatedmcrgenes (methyl coenzyme M reductase coding genes) increased when redox conditions shifted from Fe-reducing to sulfate-reducing conditions. The cytochrome genes detected were primarily fromGeobactersp. and decreased with lower subsurface redox conditions. Statistical analysis of environmental parameters and functional genes indicated that acetate, U(VI), and redox potential (Eh) were the most significant geochemical variables linked to microbial functional gene structures, and changes in microbial functional diversity were strongly related to the dominant terminal electron-accepting process following acetate addition. The study indicates that the microbial functional genes clearly reflect thein situredox conditions and the dominant microbial processes, which in turn influence uranium bioreduction. Microbial functional genes thus could be very useful for tracking microbial community structure and dynamics during bioremediation.

Metagenomics of Marine Actinomycetes: From Functional Gene Diversity to Biodiscovery

Marine OMICS ◽  
2016 ◽  
pp. 165-186 ◽  
Author(s):  
Sergey B. Zotchev ◽  
Olga N. Sekurova ◽  
D. İpek Kurtböke
Keyword(s):  
Gene Diversity ◽  
Functional Gene ◽  
Marine Actinomycetes ◽  
Functional Gene Diversity

scholarly journals Structure of recombinants from conjugational crosses between Escherichia coli donor and mismatch-repair deficient Salmonella typhimurium recipients.

Genetics ◽  
1994 ◽  
Vol 136 (1) ◽  
pp. 17-26
Author(s):  
I Matic ◽  
M Radman ◽  
C Rayssiguier
Keyword(s):  
Escherichia Coli ◽  
Salmonella Typhimurium ◽  
Mismatch Repair ◽  
Genomic Dna ◽  
Sequence Divergence ◽  
Physical Analysis ◽  
Repair System ◽  
Donor Genome ◽  
E Coli ◽  
Recombination Pathway

Abstract To get more insight into the control of homologous recombination between diverged DNA by the Mut proteins of the long-patch mismatch repair system, we have studied interspecies Escherichia coli/Salmonella typhimurium recombination. Knowing that the same recombination pathway (RecABCD) is responsible for intraspecies and interspecies recombination, we have now studied the structure (replacement vs. addition-type or other rearrangement-type recombinants) of 81 interspecies recombinants obtained in conjugational crosses between E. coli donor and mutL, mutS, mutH, mutU or mut+ S. typhimurium recipients. Taking advantage of high interspecies sequence divergence, a physical analysis was performed on one third of the E. coli Hfr genome, which was expected to be transferred to S. typhimurium F- recipients during 40 min before interruption of the mating. Probes specific for each species were hybridized on dot blots of genomic DNA, or on colonies, and the composition of the rrn operons was determined from purified genomic DNA. With very few exceptions, the structure of these interspecies recombinants corresponds to replacements of one continuous block of the recipient genome by the corresponding region of the donor genome.

scholarly journals On the evolution and physiology of cable bacteria

2019 ◽  
Vol 116 (38) ◽  
pp. 19116-19125 ◽  
Author(s):  
Kasper U. Kjeldsen ◽  
Lars Schreiber ◽  
Casper A. Thorup ◽  
Thomas Boesen ◽  
Jesper T. Bjerg ◽  
...  
Keyword(s):  
Electron Flow ◽  
Sulfide Oxidation ◽  
Gene Families ◽  
Last Common Ancestor ◽  
Long Distance ◽  
Microbial World ◽  
Single Filament ◽  
Oxic Zone ◽  
Pure Cultures ◽  
Unique Genes

Cable bacteria of the family Desulfobulbaceae form centimeter-long filaments comprising thousands of cells. They occur worldwide in the surface of aquatic sediments, where they connect sulfide oxidation with oxygen or nitrate reduction via long-distance electron transport. In the absence of pure cultures, we used single-filament genomics and metagenomics to retrieve draft genomes of 3 marine Candidatus Electrothrix and 1 freshwater Ca. Electronema species. These genomes contain >50% unknown genes but still share their core genomic makeup with sulfate-reducing and sulfur-disproportionating Desulfobulbaceae, with few core genes lost and 212 unique genes (from 197 gene families) conserved among cable bacteria. Last common ancestor analysis indicates gene divergence and lateral gene transfer as equally important origins of these unique genes. With support from metaproteomics of a Ca. Electronema enrichment, the genomes suggest that cable bacteria oxidize sulfide by reversing the canonical sulfate reduction pathway and fix CO2 using the Wood–Ljungdahl pathway. Cable bacteria show limited organotrophic potential, may assimilate smaller organic acids and alcohols, fix N2, and synthesize polyphosphates and polyglucose as storage compounds; several of these traits were confirmed by cell-level experimental analyses. We propose a model for electron flow from sulfide to oxygen that involves periplasmic cytochromes, yet-unidentified conductive periplasmic fibers, and periplasmic oxygen reduction. This model proposes that an active cable bacterium gains energy in the anodic, sulfide-oxidizing cells, whereas cells in the oxic zone flare off electrons through intense cathodic oxygen respiration without energy conservation; this peculiar form of multicellularity seems unparalleled in the microbial world.

scholarly journals Gene Families, Epistasis and the Amino Acid Preferences of Protein Homologs

2019 ◽  
Vol 15 ◽  
pp. 117693431987048
Author(s):  
Evandro Ferrada
Keyword(s):  
Amino Acid ◽  
Genetic Background ◽  
Sequence Divergence ◽  
Gene Families ◽  
Structure And Function ◽  
Sequence Evolution ◽  
Systematic Analysis ◽  
Fitness Effects ◽  
Computational Work ◽  
And Function

In order to preserve structure and function, proteins tend to preferentially conserve amino acids at particular sites along the sequence. Because mutations can affect structure and function, the question arises whether the preference of a protein site for a particular amino acid varies between protein homologs, and to what extent that variation depends on sequence divergence. Answering these questions can help in the development of models of sequence evolution, as well as provide insights on the dependence of the fitness effects of mutations on the genetic background of sequences, a phenomenon known as epistasis. Here, I comment on recent computational work providing a systematic analysis of the extent to which the amino acid preferences of proteins depend on the background mutations of protein homologs.

scholarly journals Plant sterol biosynthesis: identification of two distinct families of sterol 4alpha-methyl oxidases

2004 ◽  
Vol 378 (3) ◽  
pp. 889-898 ◽  
Author(s):  
Sylvain DARNET ◽  
Alain RAHIER
Keyword(s):  
Viral Vector ◽  
Gene Families ◽  
Amino Acid Sequences ◽  
Virus Induced Gene Silencing ◽  
Qualitative And Quantitative ◽  
Sequence Identity ◽  
Photosynthetic Eukaryotes ◽  
Membrane Bound ◽  
Haem Iron ◽  
Cdna Fragments

In plants, the conversion of cycloartenol into functional phytosterols requires the removal of the two methyl groups at C-4 by an enzymic complex including a sterol 4α-methyl oxidase (SMO). We report the cloning of candidate genes for SMOs in Arabidopsis thaliana, belonging to two distinct families termed SMO1 and SMO2 and containing three and two isoforms respectively. SMO1 and SMO2 shared low sequence identity with each other and were orthologous to the ERG25 gene from Saccharomyces cerevisiae which encodes the SMO. The plant SMO amino acid sequences possess all the three histidine-rich motifs (HX3H, HX2HH and HX2HH), characteristic of the small family of membrane-bound non-haem iron oxygenases that are involved in lipid oxidation. To elucidate the precise functions of SMO1 and SMO2 gene families, we have reduced their expression by using a VIGS (virus-induced gene silencing) approach in Nicotiana benthamiana. SMO1 and SMO2 cDNA fragments were inserted into a viral vector and N. benthamiana inoculated with the viral transcripts. After silencing with SMO1, a substantial accumulation of 4,4-dimethyl-9β,19-cyclopropylsterols (i.e. 24-methylenecycloartanol) was obtained, whereas qualitative and quantitative levels of 4α-methylsterols were not affected. In the case of silencing with SMO2, a large accumulation of 4α-methyl-Δ7-sterols (i.e. 24-ethylidenelophenol and 24-ethyllophenol) was found, with no change in the levels of 4,4-dimethylsterols. These clear and distinct biochemical phenotypes demonstrate that, in contrast with animals and fungi, in photosynthetic eukaryotes, these two novel families of cDNAs are coding two distinct types of C-4-methylsterol oxidases controlling the level of 4,4-dimethylsterol and 4α-methylsterol precursors respectively.

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