The Translation Start Codon Region Is Sensitive to Antisense PNA Inhibition inEscherichia coli

2003 ◽  
Vol 13 (6) ◽  
pp. 427-433 ◽  
Author(s):  
Rikard Dryselius ◽  
Satish Kumar Aswasti ◽  
Gunaratna K. Rajarao ◽  
Peter E. Nielsen ◽  
Liam Good
Keyword(s):  
Start Codon ◽  
Inescherichia Coli ◽  
Translation Start Codon ◽  
Translation Start

The human connexin32 gene is transcribed from two tissue-specific promoters

1996 ◽  
Vol 16 (3) ◽  
pp. 239-248 ◽  
Author(s):  
Isaac M. Neuhaus ◽  
Linda Bone ◽  
Suping Wang ◽  
Victor Ionasescu ◽  
Rudolf Werner
Keyword(s):  
Start Codon ◽  
Coding Region ◽  
Translation Start Codon ◽  
Tissue Specific ◽  
Gene Coding ◽  
Large Intron ◽  
Translation Start ◽  
Junction Protein ◽  
Liver And Pancreas ◽  
Alternatively Spliced

The connexin32 (cx32) gene codes for the gap junction protein found in liver, pancreas and nervous tissue. Recently mutations in the coding region of this gene have been associated with the dominant X-linked form of Charcot-Marie-Tooth (CMTX1) neuropathy. Since some CMTX1 patients show no mutations in their cx32 gene coding region, it was speculated that these patients carry mutations in the promoter region of the gene. This paper describes the organization of the human cx32 gene and its tissue-specific transcription. The gene consists of three exons that are alternatively spliced to produce mRNAs with different 5′-untranslated regions (UTRs). Transcription is initiated from two tissue-specific promoters. In liver and pancreas, promoter P1. located more than 8 kb upstream of the translation start codon, is used, and the transcript is processed to remove a large intron. In contrast, in nerve cells, transcription is initiated from promoter P2, located 497 bp upstream from the translation start codon, and the transcript is processed to remove a small 355-pb intron. The downstream exon. which includes the entire coding sequence, is shared by both mRNAs. CMTX1 patients with a normal cx32 coding region are expected to have mutations in this newly described promoter P2 rather than the known promoter P1.

scholarly journals Functional Analyses of the Promoters in the Lantibiotic Mutacin II Biosynthetic Locus in Streptococcus mutans

1999 ◽  
Vol 65 (2) ◽  
pp. 652-658 ◽  
Author(s):  
Fengxia Qi ◽  
Ping Chen ◽  
Page W. Caufield
Keyword(s):  
Gene Expression ◽  
Complex System ◽  
Streptococcus Mutans ◽  
Promoter Activity ◽  
Growth Stage ◽  
Start Codon ◽  
Early Stationary Phase ◽  
Translation Start Codon ◽  
Translation Start ◽  
Functional Analyses

ABSTRACT The lantibiotic bacteriocin mutacin II is produced by the group IIStreptococcus mutans. The mutacin II biosynthetic locus consists of seven genes, mutR, -A, -M, -T, -F, -E, and -G, organized as two operons. The mutAMTFEGoperon is transcribed from the mutA promoter 55 bp upstream of the translation start codon for MutA, while the mutRpromoter is 76 bp upstream of the mutR structural gene. Expression of the mutA promoter is regulated by the components of the growth medium, while the mutR promoter activity does not seem to be affected by these conditions. Inactivation of mutR abolishes transcription of the mutAoperon but does not affect its own promoter activity. The expressions of both mutA and mutR promoters are independent of the growth stage, while the production of mutacin II is only elevated at the early stationary phase. Taken together, these results suggest that expression of the mutacin operon is regulated by a complex system involving transcriptional and posttranscriptional or posttranslational controls.

Genetic and physical analysis of the chicken tk gene

1984 ◽  
Vol 4 (9) ◽  
pp. 1769-1776
Author(s):  
G F Merrill ◽  
R M Harland ◽  
M Groudine ◽  
S L McKnight
Keyword(s):  
Start Codon ◽  
Nucleotide Sequencing ◽  
Deletion Mapping ◽  
Physical Analysis ◽  
Translation Start Codon ◽  
Kinase Gene ◽  
Cdna Sequences ◽  
Intervening Sequences ◽  
Translation Start ◽  
Functional Boundary

Several aspects of the structure of the chicken thymidine kinase gene (tk) have been resolved as a result of genetic experiments and nucleotide sequencing. Deletion mapping established the locations of two functional boundaries in a region thought to correspond to the 5' terminus of the gene. One such boundary coincides with a transcriptional promoter, and the other coincides with the translation start codon of the chicken tk polypeptide. Similar deletion mapping assays identified a functional boundary at the 3' terminus of the gene. DNA sequence analysis confirms the prediction that this 3' region encodes the carboxyl terminus of the tk polypeptide. A recombinant cDNA clone complementary to genomic tk sequences was isolated. A comparison between genomic and cDNA sequences reveals the locations of six intervening sequences and allows prediction of the complete amino acid sequence of the chicken tk polypeptide.

scholarly journals Efficient use of a translation start codon in BDNF exon I

2015 ◽  
Vol 134 (6) ◽  
pp. 1015-1025 ◽  
Author(s):  
Indrek Koppel ◽  
Jürgen Tuvikene ◽  
Ingrid Lekk ◽  
Tõnis Timmusk
Keyword(s):  
Start Codon ◽  
Translation Start Codon ◽  
Translation Start

scholarly journals Aquareovirus Effects Syncytiogenesis by Using a Novel Member of the FAST Protein Family Translated from a Noncanonical Translation Start Site

2009 ◽  
Vol 83 (11) ◽  
pp. 5951-5955 ◽  
Author(s):  
Trina Racine ◽  
Tara Hurst ◽  
Chris Barry ◽  
Jingyun Shou ◽  
Frederick Kibenge ◽  
...  
Keyword(s):  
Syncytium Formation ◽  
Integral Membrane Protein ◽  
Genome Segment ◽  
Start Codon ◽  
Start Site ◽  
Translation Start Site ◽  
Translation Start Codon ◽  
Translation Start ◽  
Fast Proteins ◽  
Distinct Member

ABSTRACT As nonenveloped viruses, the aquareoviruses and orthoreoviruses are unusual in their ability to induce cell-cell fusion and syncytium formation. While an extraordinary family of fusion-associated small transmembrane (FAST) proteins is responsible for orthoreovirus syncytiogenesis, the basis for aquareovirus-induced syncytiogenesis is unknown. We now report that the S7 genome segment of an Atlantic salmon reovirus is polycistronic and uses a noncanonical CUG translation start codon to produce a 22-kDa integral membrane protein responsible for syncytiogenesis. The aquareovirus p22 protein represents a fourth distinct member of the FAST family with a unique repertoire and arrangement of structural motifs.

scholarly journals Promoter Characterization and Constitutive Expression of the Escherichia coli gcvR Gene

1998 ◽  
Vol 180 (7) ◽  
pp. 1803-1807 ◽  
Author(s):  
Angela C. Ghrist ◽  
George V. Stauffer
Keyword(s):  
Escherichia Coli ◽  
Transcription Start Site ◽  
Promoter Region ◽  
Start Codon ◽  
Start Site ◽  
Transcription Start ◽  
Translation Start Codon ◽  
Translation Start ◽  
Glycine Cleavage ◽  
Extension Analysis

ABSTRACT The Escherichia coli glycine cleavage repressor protein (GcvR) negatively regulates expression of the glycine cleavage operon (gcv). In this study, the gcvR translational start site was determined by N-terminal amino acid sequence analysis of a GcvR-LacZ fusion protein. Primer extension analysis of thegcvR promoter region identified a primary transcription start site 27 bp upstream of the UUG translation start site and a minor transcription start site approximately 100 bp upstream of the translation start codon. The -10 and -35 promoter regions upstream of the primary transcription start site were defined by mutational analysis. Expression of a gcvR-lacZ fusion was unaltered in the presence of glycine or inosine, molecules known to induce or repress expression of gcv, respectively. In addition, it was shown that gcvR-lacZ expression is neither regulated by the glycine cleavage activator protein (GcvA) nor autogenously regulated by GcvR. From DNA sequence analysis, it was predicted that the translation start codon of the downstream bcp gene overlaps the gcvR stop codon, suggesting that these genes may form an operon. However, a down mutation in the -10 promoter region of gcvR had no effect on the expression of a downstreambcp-lacZ fusion, and primer extension analysis of thebcp promoter region demonstrated that bcp has its own promoter within the gcvR coding sequence. These results show that gcvR and bcp do not form an operon. Furthermore, the deletion of bcp from the chromosome had no effect on gcv-lacZ expression.

30 A TRANSCRIPTION ACTIVATOR-LIKE EFFECTOR NUCLEASE (TALEN)-MEDIATED UNIVERSAL GENE KNOCK-IN STRATEGY FOR MAMMARY GLANDS-SPECIFIC EXPRESSION OF RECOMBINANT PROTEINS IN DAIRY CATTLE

2014 ◽  
Vol 26 (1) ◽  
pp. 129 ◽  
Author(s):  
S. Lee ◽  
H. Park ◽  
I. Kong ◽  
Z. Wang
Keyword(s):  
Dairy Cattle ◽  
Single Cell ◽  
Recombinant Proteins ◽  
Mammary Glands ◽  
Start Codon ◽  
Regulatory Sequences ◽  
Transcription Activator ◽  
Translation Start Codon ◽  
Casein Gene ◽  
Translation Start

To harness the great capability of producing biologically active recombinant proteins with animal mammary glands, active research has been carried out in the past several decades to develop transgenic animals as bioreactors. However, when a transgene is introduced in the animal genome by random integration, the transgene tends to be subjected to epigenetic silencing, due to the so-called position effect from the chromatin environments surrounding the transgene integration sites, thereby resulting in low-level expression or total suppression. We report a universal transgenic strategy to knock in (KI) transgenes into the bovine β-casein gene locus allowing the expression of a transgene to be totally under the control of the endogenous regulatory sequences of the bovine β-casein gene. This universal KI strategy comprises two key components: one is the design of transcription activator-like effector nuclease (TALEN) constructs targeting the start codon region of bovine β-casein gene, and the other is the design of KI vectors in which a transgene of choice is flanked with homologous arms isolated from the ~500-bp bovine genomic DNAs immediately 5′ and 3′, respectively, of the translation start codon of the bovine β-casein gene. By using the human erythropoietin (hEPO) as the model transgene, we demonstrated that a transgene can be highly efficiently integrated immediately after the translation start codon of the bovine β-casein gene. In brief, the TALEN constructs were assembled by using the Golden Gate protocol. To KI the hEPO transgene, early passage (<5) of fibroblasts established from Holstein dairy cattle were cultured into full confluence in Dulbecco's modified Eagle medium (DMEM) supplemented with 10% fetal bovine serum (FBS), harvested with 0.25% trypsin-EDTA, and co-transfected with KI vector and the TALEN constructs by the Amaxa Nucleofector system. For each experiment, 106 cells were transfected with 5 μg of KI vector and 5 μg of TALEN constructs. After 72 h post-transfection, cells were harvested and subjected to limiting dilution to obtain single-cell derived colonies. To screen for single-cell derived colonies carrying the correctly KI of hEPO in the β-casein locus, we performed genomic PCR amplifying the genomic junctions created by the KI of hEPO gene into the bovine genome. We identified and established 2 hEPO transgenic bovine fibroblast cell lines after screening 10 single-cell derived colonies from the transfected cells (20%). The genotype of these 2 colonies was also confirmed by sequencing the PCR products. We have initiated the effort to produce hEPO transgenic cattle by somatic cell nuclear transfer (SCNT), and the animal cloning results will be reported at the conference.

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