scholarly journals Three genes for the elongation factor EF-1 alpha in Mucor racemosus.

1986 ◽  
Vol 6 (2) ◽  
pp. 593-600 ◽  
Author(s):  
J E Linz ◽  
C Katayama ◽  
P S Sypherd
Keyword(s):  
Nucleotide Sequence ◽  
Genomic Dna ◽  
Sequence Data ◽  
Elongation Factor ◽  
Dna Fragments ◽  
Mucor Racemosus ◽  
S1 Nuclease ◽  
Nucleotide Sequence Data ◽  
Flanking Regions ◽  
Overlapping Region

We cloned three genes from Mucor racemosus coding for protein synthesis elongation factor 1 alpha (EF-1 alpha). A 110-base-pair (bp) EF-1 alpha-specific cDNA clone was identified by hybrid-selected translation. The nucleotide sequence of the cDNA showed significant homology to a region of the Saccharomyces cerevisiae genes for EF-1 alpha (TEF1 and TEF2). The cDNA was used to isolate an 850-bp EcoRI genomic DNA fragment containing a portion of the EF-1 alpha gene. Screening of a lambda/M. racemosus genomic DNA bank with the 850-bp EcoRI probe resulted in the identification of three DNA fragments containing a common 850-bp EcoRI fragment within a short overlapping region. S1 nuclease analysis of the three EF-1 alpha DNA fragments showed that the EF-1 alpha transcript covered the short overlapping region in the clones. Restriction fragments purified from flanking regions in each clone were used to probe a HindIII digest of M. racemosus genomic DNA. Each flanking probe hybridized to one of three DNA fragments which hybridized to the 850-bp EF-1 alpha-specific probe. Nucleotide sequence data from two random "shotgun clones" of one of the three genes show good homology to two regions of S. cerevisiae TEF1. The data indicate the presence of three genes for EF-1 alpha in M. racemosus located at unique sites in the genome.

Three genes for the elongation factor EF-1 alpha in Mucor racemosus

1986 ◽  
Vol 6 (2) ◽  
pp. 593-600
Author(s):  
J E Linz ◽  
C Katayama ◽  
P S Sypherd
Keyword(s):  
Nucleotide Sequence ◽  
Genomic Dna ◽  
Sequence Data ◽  
Elongation Factor ◽  
Dna Fragments ◽  
Mucor Racemosus ◽  
S1 Nuclease ◽  
Nucleotide Sequence Data ◽  
Flanking Regions ◽  
Overlapping Region

We cloned three genes from Mucor racemosus coding for protein synthesis elongation factor 1 alpha (EF-1 alpha). A 110-base-pair (bp) EF-1 alpha-specific cDNA clone was identified by hybrid-selected translation. The nucleotide sequence of the cDNA showed significant homology to a region of the Saccharomyces cerevisiae genes for EF-1 alpha (TEF1 and TEF2). The cDNA was used to isolate an 850-bp EcoRI genomic DNA fragment containing a portion of the EF-1 alpha gene. Screening of a lambda/M. racemosus genomic DNA bank with the 850-bp EcoRI probe resulted in the identification of three DNA fragments containing a common 850-bp EcoRI fragment within a short overlapping region. S1 nuclease analysis of the three EF-1 alpha DNA fragments showed that the EF-1 alpha transcript covered the short overlapping region in the clones. Restriction fragments purified from flanking regions in each clone were used to probe a HindIII digest of M. racemosus genomic DNA. Each flanking probe hybridized to one of three DNA fragments which hybridized to the 850-bp EF-1 alpha-specific probe. Nucleotide sequence data from two random "shotgun clones" of one of the three genes show good homology to two regions of S. cerevisiae TEF1. The data indicate the presence of three genes for EF-1 alpha in M. racemosus located at unique sites in the genome.

Expression of three genes for elongation factor 1 alpha during morphogenesis of Mucor racemosus

1987 ◽  
Vol 7 (5) ◽  
pp. 1925-1932
Author(s):  
J E Linz ◽  
P S Sypherd
Keyword(s):  
Sequence Data ◽  
Elongation Factor ◽  
Yeast Cells ◽  
Northern Analysis ◽  
Mucor Racemosus ◽  
S1 Nuclease ◽  
Nucleotide Sequence Data ◽  
Elongation Factor 1 Alpha ◽  
Nuclease Mapping ◽  
Elongation Factor 1

Three genes, TEF-1, -2, and -3, encode elongation factor 1 alpha in Mucor racemosus. Neutral and alkaline S1 nuclease analyses revealed that the genetic organization is unique for each of the genes. The number and size of the intervening sequences vary in these closely related genes, which suggests that complex genetic rearrangements gave rise to the elongation factor 1 alpha gene family. Nucleotide sequence data from restriction fragments isolated from the 5' and 3' ends of TEF-2 and -3 confirmed the presence of a second intervening sequence in these genes. These data along with S1 nuclease mapping revealed a region at the 3' end of the three genes which was predicted to be transcribed but untranslated. Unique oligonucleotides containing 19 bases were synthesized to hybridize to this unique trailer region in the elongation factor 1 alpha transcripts. These oligonucleotides were used as probes in standard Northern analysis of RNA purified from M. racemosus cells of several morphological types. It was determined that all three genes were expressed in the cell morphological types studied. However, the accumulated level of transcript derived from each gene varied considerably, with TEF-1 mRNA present in approximately twofold greater quantity than the TEF-3 transcript and up to sixfold greater quantity than TEF-2. The level of TEF-1 and -2 mRNA varied little among the cell morphological types studied, whereas TEF-3 mRNA was present in twofold greater quantity in sporangiospores than in either germlings or yeast cells which had been induced to undergo morphogenesis to hyphae. These data suggest that there is differential expression of the genes encoding elongation factor 1 alpha in M. racemosus. At least one gene, TEF-3, shows a morphology-specific pattern of transcript accumulation.

scholarly journals Expression of three genes for elongation factor 1 alpha during morphogenesis of Mucor racemosus.

1987 ◽  
Vol 7 (5) ◽  
pp. 1925-1932 ◽  
Author(s):  
J E Linz ◽  
P S Sypherd
Keyword(s):  
Sequence Data ◽  
Elongation Factor ◽  
Yeast Cells ◽  
Northern Analysis ◽  
Mucor Racemosus ◽  
S1 Nuclease ◽  
Nucleotide Sequence Data ◽  
Elongation Factor 1 Alpha ◽  
Nuclease Mapping ◽  
Elongation Factor 1

Three genes, TEF-1, -2, and -3, encode elongation factor 1 alpha in Mucor racemosus. Neutral and alkaline S1 nuclease analyses revealed that the genetic organization is unique for each of the genes. The number and size of the intervening sequences vary in these closely related genes, which suggests that complex genetic rearrangements gave rise to the elongation factor 1 alpha gene family. Nucleotide sequence data from restriction fragments isolated from the 5' and 3' ends of TEF-2 and -3 confirmed the presence of a second intervening sequence in these genes. These data along with S1 nuclease mapping revealed a region at the 3' end of the three genes which was predicted to be transcribed but untranslated. Unique oligonucleotides containing 19 bases were synthesized to hybridize to this unique trailer region in the elongation factor 1 alpha transcripts. These oligonucleotides were used as probes in standard Northern analysis of RNA purified from M. racemosus cells of several morphological types. It was determined that all three genes were expressed in the cell morphological types studied. However, the accumulated level of transcript derived from each gene varied considerably, with TEF-1 mRNA present in approximately twofold greater quantity than the TEF-3 transcript and up to sixfold greater quantity than TEF-2. The level of TEF-1 and -2 mRNA varied little among the cell morphological types studied, whereas TEF-3 mRNA was present in twofold greater quantity in sporangiospores than in either germlings or yeast cells which had been induced to undergo morphogenesis to hyphae. These data suggest that there is differential expression of the genes encoding elongation factor 1 alpha in M. racemosus. At least one gene, TEF-3, shows a morphology-specific pattern of transcript accumulation.

scholarly journals Characterization of squalene synthase gene from Gymnema sylvestre R. Br.

2021 ◽  
Vol 10 (1) ◽  
Author(s):  
Kuldeepsingh A. Kalariya ◽  
Ram Prasnna Meena ◽  
Lipi Poojara ◽  
Deepa Shahi ◽  
Sandip Patel
Keyword(s):  
Dna Sequences ◽  
Genomic Dna ◽  
Competitive Inhibition ◽  
Sequence Data ◽  
Homology Model ◽  
Squalene Synthase ◽  
Gymnema Sylvestre ◽  
Gardenia Jasminoides ◽  
Ramachandran Plots ◽  
Flanking Regions

Abstract Background Squalene synthase (SQS) is a rate-limiting enzyme necessary to produce pentacyclic triterpenes in plants. It is an important enzyme producing squalene molecules required to run steroidal and triterpenoid biosynthesis pathways working in competitive inhibition mode. Reports are available on information pertaining to SQS gene in several plants, but detailed information on SQS gene in Gymnema sylvestre R. Br. is not available. G. sylvestre is a priceless rare vine of central eco-region known for its medicinally important triterpenoids. Our work aims to characterize the GS-SQS gene in this high-value medicinal plant. Results Coding DNA sequences (CDS) with 1245 bp length representing GS-SQS gene predicted from transcriptome data in G. sylvestre was used for further characterization. The SWISS protein structure modeled for the GS-SQS amino acid sequence data had MolProbity Score of 1.44 and the Clash Score 3.86. The quality estimates and statistical score of Ramachandran plots analysis indicated that the homology model was reliable. For full-length amplification of the gene, primers designed from flanking regions of CDS encoding GS-SQS were used to get amplification against genomic DNA as template which resulted in approximately 6.2-kb sized single-band product. The sequencing of this product through NGS was carried out generating 2.32 Gb data and 3347 number of scaffolds with N50 value of 457 bp. These scaffolds were compared to identify similarity with other SQS genes as well as the GS-SQSs of the transcriptome. Scaffold_3347 representing the GS-SQS gene harbored two introns of 101 and 164 bp size. Both these intronic regions were validated by primers designed from adjoining outside regions of the introns on the scaffold representing GS-SQS gene. The amplification took place when the template was genomic DNA and failed when the template was cDNA confirmed the presence of two introns in GS-SQS gene in Gymnema sylvestre R. Br. Conclusion This study shows GS-SQS gene was very closely related to Coffea arabica and Gardenia jasminoides and this gene harbored two introns of 101 and 164 bp size.

ON THE RATIONALE AND UTILITY OF WEIGHTING NUCLEOTIDE SEQUENCE DATA

Cladistics ◽  
1992 ◽  
Vol 8 (1) ◽  
pp. 73-83 ◽  
Author(s):  
Victor A. Albert ◽  
Brent D. Mishler
Keyword(s):  
Nucleotide Sequence ◽  
Sequence Data ◽  
Nucleotide Sequence Data

A new method for RAPD primers selection based on primer bias in nucleotide sequence data

2006 ◽  
Vol 126 (4) ◽  
pp. 415-423 ◽  
Author(s):  
J.J. Li ◽  
G.L. Pei ◽  
H.X. Pang ◽  
A. Bilderbeck ◽  
S.S. Chen ◽  
...  
Keyword(s):  
Nucleotide Sequence ◽  
Sequence Data ◽  
New Method ◽  
Nucleotide Sequence Data
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