Molecular cloning and nucleotide sequence analysis of the putative cDNA for the precursor molecule of the chicken LH-β subunit

1989 ◽  
Vol 3 (2) ◽  
pp. 129-137 ◽  
Author(s):  
T. Noce ◽  
H. Ando ◽  
T. Ueda ◽  
K. Kubokawa ◽  
T. Higashinakagawa ◽  
...  
Keyword(s):  
Nucleotide Sequence ◽  
Cdna Clone ◽  
Rabbit Antiserum ◽  
Nucleotide Sequence Analysis ◽  
Cdna Expression Library ◽  
Β Subunit ◽  
Coding Region ◽  
Precursor Molecule ◽  
Cdna Insert ◽  
Hybridization Probe

ABSTRACT A cDNA expression library was constructed from poly(A)+ RNA of broiler chicken adenohypophyses using λ gt11 as a vector. After screening with a rabbit antiserum against chicken LH, a cDNA clone (L12) containing a 436 bp insert was obtained. Using a subclone of L12 in pUC19 (pL12) as the hybridization probe, another cDNA clone (LF127) with a 533 bp insert was isolated. The LF127 contained the full-length cDNA encoding the putative chicken LH-β subunit precursor molecule. Hybridization of the pL12 cDNA insert to adenohypophysial RNA showed that chicken and Japanese quail adenohypophyses contained RNA species of about 0·8 and 1·0 kb respectively. The amount of this RNA species was ten times higher in adult male quails kept under long days at room temperature than in those kept under short days at 7 °C. In-situ hybridization experiments showed the exclusive distribution of the signal in the LH cells of the adenohypophysis. The similarity of the nucleotide sequence of the apoprotein-coding region of LH-β cDNA of the chicken to that of mammals is lower than that among mammals. The deduced amino acid sequence of the chicken LH-β subunit supports the hypothesis that the number of proline residues increases in the LH-β subunit the closer phylogenetically the vertebrate is to mammals.

Molecular cloning of a rat uterine gap junction protein and analysis of gene expression during gestation

1991 ◽  
Vol 260 (5) ◽  
pp. E787-E793 ◽  
Author(s):  
L. M. Lang ◽  
E. C. Beyer ◽  
A. L. Schwartz ◽  
J. D. Gitlin
Keyword(s):  
Gap Junctions ◽  
Gap Junction ◽  
Cdna Clone ◽  
Blot Analysis ◽  
Nucleotide Sequence Analysis ◽  
Uterine Tissue ◽  
Coding Region ◽  
Junction Protein ◽  
Gap Junction Protein ◽  
Rna Blot Analysis

To study the molecular mechanisms controlling the rapid increase in myometrial gap junctions observed in the parturient uterus, we have isolated a full-length cDNA clone corresponding to a rat uterine gap junction protein. Nucleotide sequence analysis of the cDNA clone reveals complete identity of the coding region with that of a previously reported heart gap junction protein (connexin43). Southern blot analysis suggests that the gene encoding this gap junction protein exists as a single copy in the rat haploid genome and contains no introns within the coding region. RNA blot analysis with this gap junction cDNA reveals a single 3.0-kb mRNA in uterine tissue without changes in transcript size throughout gestation. When normalized to the amount of 28S rRNA, the relative abundance of the connexin43 transcript in uterine tissue is quite constant between the nonpregnant state, during gestation, intrapartum, and postpartum. Similar size transcripts are shown by RNA blot analysis to be present in heart, lung, liver, brain, and skeletal muscle, and these transcripts are identified by the same 3'-nontranslated sequence probe. The results of these studies suggest that rat connexin43 is encoded by a single gene that is transcribed to identical transcripts in heart, uterus, and other tissues. They further suggest that changes in the abundance of connexin43 transcript are unlikely to be responsible for the abrupt increase in connexin43-containing myometrial gap junctions at term.

scholarly journals Analysis of a New Human Parechovirus Allows the Definition of Parechovirus Types and the Identification of RNA Structural Domains

2007 ◽  
Vol 81 (2) ◽  
pp. 1013-1021 ◽  
Author(s):  
Mohammed Al-Sunaidi ◽  
Çiğdem H. Williams ◽  
Pamela J. Hughes ◽  
David P. Schnurr ◽  
Glyn Stanway
Keyword(s):  
Nucleotide Sequence ◽  
Untranslated Region ◽  
Molecular Techniques ◽  
Nucleotide Sequence Analysis ◽  
Important Criterion ◽  
Coding Region ◽  
Human Parechovirus ◽  
Structural Domains ◽  
Genetic Groups ◽  
Definition Of

ABSTRACT Human parechoviruses (HPeV), members of the Parechovirus genus of Picornaviridae, are frequent pathogens but have been comparatively poorly studied, and little is known of their diversity, evolution, and molecular biology. To increase the amount of information available, we have analyzed 7 HPeV strains isolated in California between 1973 and 1992. We found that, on the basis of VP1 sequences, these fall into two genetic groups, one of which has not been previously observed, bringing the number of known groups to five. While these correlate partly with the three known serotypes, two members of the HPeV2 serotype belong to different genetic groups. In view of the growing importance of molecular techniques in diagnosis, we suggest that genotype is an important criterion for identifying viruses, and we propose that the genetic groups we have defined should be termed human parechovirus types 1 to 5. Complete nucleotide sequence analysis of two of the Californian isolates, representing two types, confirmed the identification of a new genetic group and suggested a role for recombination in parechovirus evolution. It also allowed the identification of a putative HPeV1 cis-acting replication element, which is located in the VP0 coding region, as well as the refinement of previously predicted 5′ and 3′ untranslated region structures. Thus, the results have significantly improved our understanding of these common pathogens.

scholarly journals A low polymorphic mouse H-2 class I gene from the Tla complex is expressed in a broad variety of cell types.

1987 ◽  
Vol 166 (2) ◽  
pp. 341-361 ◽  
Author(s):  
C Transy ◽  
S R Nash ◽  
B David-Watine ◽  
M Cochet ◽  
S W Hunt ◽  
...  
Keyword(s):  
Nucleotide Sequence ◽  
Cdna Clone ◽  
Cell Types ◽  
Class I ◽  
Surface Molecule ◽  
Coding Region ◽  
Genes Encoding ◽  
Broad Variety ◽  
I Gene

We have previously described the isolation of pH-2d-37, a cDNA clone that encodes a so far unknown, poorly polymorphic, class I surface molecule. We report here the isolation of the corresponding gene, its nucleotide sequence, and its localization in the Tla region of the murine MHC. Using a RNase mapping assay, we have confirmed that the second domain coding region of the 37 gene displays very limited polymorphism, and that the gene is transcribed in a broad variety of cell types, in contrast to the genes encoding the known Qa and TL antigens. Possible functions are discussed.

The nucleotide sequence of a cDNA clone containing the entire coding region for mouse X-chromo- some-linked phosphoglycerate kinase

Gene ◽  
1986 ◽  
Vol 45 (3) ◽  
pp. 275-280 ◽  
Author(s):  
Mori Nozomu ◽  
Judith Singer-Sam ◽  
Lee Chi-Yu ◽  
Arthur D. Riggs
Keyword(s):  
Nucleotide Sequence ◽  
Cdna Clone ◽  
Phosphoglycerate Kinase ◽  
Coding Region

Structure and organization of the C4 genes

1984 ◽  
Vol 306 (1129) ◽  
pp. 379-388 ◽  
Keyword(s):  
Nucleotide Sequence ◽  
Cdna Clone ◽  
Cosmid Library ◽  
Null Alleles ◽  
Nucleotide Sequence Analysis ◽  
Substantial Part ◽  
Class Iii ◽  
Class Differences ◽  
Library Characterization

This 200000 M r serum protein is coded for by at least two separate loci, C4A and C4B , which map in the HLA Class III region on chromosome 6 in man. Both loci are highly polymorphic with more than 30 alleles, including null alleles assigned to the two loci. The complete nucleotide sequence of a full length C4A cDNA clone and a substantial part of a C4B cDNA clone has shown class differences which can be used to synthesize nucleotide probes specific for C4A and C4B. Three C4 loci of approximately 16 kilobases each spaced by 10 kilobases have been identified in DNA from one individual and aligned 30 kilobases from the factor B gene by overlapping cloned genomic fragments from a cosmid library. Characterization of these genes by restriction mapping, nucleotide sequence analysis and hybridization with C4A and C4B specific synthetic oligonucleotides show that these genes are very similar.

scholarly journals Isolation, Characterization, and Expression of the Gene Encoding the β Subunit of the Mitochondrial Processing Peptidase fromBlastocladiella emersonii

1998 ◽  
Vol 180 (15) ◽  
pp. 3967-3972 ◽  
Author(s):  
Cíntia Renata Costa Rocha ◽  
Suely Lopes Gomes
Keyword(s):  
Genomic Library ◽  
Nucleotide Sequence Analysis ◽  
Mrna Levels ◽  
Β Subunit ◽  
Coding Region ◽  
Gene Encoding ◽  
Western Blots ◽  
Calculated Molecular Mass ◽  
Mitochondrial Processing Peptidase ◽  
Processing Peptidase

ABSTRACT A 2.3-kb BamHI-KpnI fragment was isolated from a partial genomic library and shown by nucleotide sequence analysis to contain the entire coding region of the gene encoding the β subunit of the Blastocladiella mitochondrial processing peptidase (β-MPP). The predicted β-MPP protein has 465 amino acids and a calculated molecular mass of 50.8 kDa. S1 nuclease protection assays revealed an intron, 209 bp in size, interrupting the coding region between the putative signal sequence and the mature protein. Northern blot analysis showed that β-MPP mRNA levels decrease significantly during B. emersonii sporulation, reaching basal levels in the zoospore stage. The amount of β-MPP protein, determined in Western blots, unlike its mRNA, does not vary significantly throughout the fungal life cycle.

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