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See detailMolecular cloning and characterization of two forms of trout growth hormone cDNA: expression and secretion of tGH-II by Escherichia coli
Rentier-Delrue, Françoise ULg; Swennen, D.; Mercier, L. et al

in DNA (1989), 8(2), 109-17

We constructed a cDNA library using mRNA isolated from rainbow trout pituitaries. Two types of cDNA clones encoding growth hormone (GH) were isolated and their complete nucleotide sequences determined ... [more ▼]

We constructed a cDNA library using mRNA isolated from rainbow trout pituitaries. Two types of cDNA clones encoding growth hormone (GH) were isolated and their complete nucleotide sequences determined. Twenty seven nucleotide substitutions in the coding region and 108 in the noncoding region distinguish the cDNAs of trout GH-I and II. Both cDNAs encode polypeptides of 210 amino acids, including a putative signal peptide of 22 amino acids, which differ by 12 residues. In both trout and salmon, GH-I mRNA is predominant, which suggests that the variation in the amount of secreted GH originates from a transcriptional event. Moreover, comparison of rainbow trout and chum salmon GH reveals that, in both cases, the predominant GH-I has mutated less than its GH-II counterpart. Mature tGH-II was expressed in Escherichia coli using the pIN-III-ompA-Hind secretion vector. [less ▲]

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See detailRainbow trout prolactin cDNA cloning in Escherichia coli
Mercier, L.; Rentier-Delrue, Françoise ULg; Swennen, D. et al

in DNA (1989), 8(2), 119-25

We describe the isolation and characterization of a cDNA for trout prolactin (tPrl). An extensive analysis of tPrl recombinant clones by restriction analysis and sequencing revealed the presence of only ... [more ▼]

We describe the isolation and characterization of a cDNA for trout prolactin (tPrl). An extensive analysis of tPrl recombinant clones by restriction analysis and sequencing revealed the presence of only one form of tPrl mRNA. The deduced protein sequence consists of 210 amino acids, including a signal peptide of 23 amino acids. The amino acid sequence of the mature protein is compared among teleosts and mammals, showing two domains of strong similarity that may be involved in biological activity. [less ▲]

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See detailPituitary-specific factor binding to the human prolactin, growth hormone, and placental lactogen genes
Lemaigre, F. P.; Peers, Bernard ULg; Lafontaine, D. A. et al

in DNA (1989), 8(3), 149-59

The human genes coding for growth hormone (GH), chorionic somatomammotropin (placental lactogen, CS), and prolactin (Prl) are related evolutionarily but are expressed in phenotypically distinct cell types ... [more ▼]

The human genes coding for growth hormone (GH), chorionic somatomammotropin (placental lactogen, CS), and prolactin (Prl) are related evolutionarily but are expressed in phenotypically distinct cell types despite their nucleotide sequence homology. We show here that the promoters of the human Prl and CS genes contain cis-acting sequences that confer pituitary-specific expression in a cell-free transcription assay. Similar data are obtained with the human GH gene, consistent with earlier work by others. Footprinting analysis shows that neighboring sequences in each of these three promoters are protected from deoxyribonuclease I digestion by rat pituitary cell extracts. Footprinting competition experiments and gel retardation assays with synthetic oligonucleotides suggest that a single factor is responsible for the pituitary-specific footprints seen on the human Prl, CS, and GH genes. They also suggest that this factor is identical or closely related to the trans-acting factor GHF-1/Pit-1. Similarities with and differences from the rat GH and Prl genes are discussed. [less ▲]

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See detailTilapia prolactin: molecular cloning of two cDNAs and expression in Escherichia coli
Rentier-Delrue, Françoise ULg; Swennen, D.; Prunet, P. et al

in DNA (1989), 8(4), 261-70

We have isolated cDNA clones encoding tilapia prolactin (tiPRL) from a cDNA library prepared from tilapia (Oreochromis niloticus) anterior pituitary glands. A trout PRL cDNA fragment was used as ... [more ▼]

We have isolated cDNA clones encoding tilapia prolactin (tiPRL) from a cDNA library prepared from tilapia (Oreochromis niloticus) anterior pituitary glands. A trout PRL cDNA fragment was used as hybridization probe to select the recombinant plasmids carrying the tiPRL coding sequence. Two types of PRL cDNA were isolated and their complete nucleotide sequence determined. The larger cDNA (tiPRL-I) codes for a polypeptide of 212 amino acids, including a putative signal sequence of 24 amino acids, and contains a 3' untranslated region of 787 bp. The second prolactin cDNA (tiPRL-II) encodes a polypeptide of 200 amino acids, including a presumptive signal peptide of 23 amino acids, and contains a noncoding region of 512 bp. tiPRL-I and tiPRL-II cDNA sequences are 81% similar, whereas the encoded proteins share 69% amino acid identity at optimal alignment. Mature tiPRL-I was efficiently expressed in Escherichia coli carrying a plasmid in which the tiPRL-I cDNA was under the control of the phi 10 promoter of T7 bacteriophage. The new recombinant protein representing about 45% of the total cellular proteins was found in inclusion bodies and cross-reacted with salmon PRL antiserum. [less ▲]

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See detailTilapia growth hormone: molecular cloning of cDNA and expression in Escherichia coli
Rentier-Delrue, Françoise ULg; Swennen, D.; Philippart, Jean-Claude ULg et al

in DNA (1989), 8(4), 271-8

A cDNA library was prepared from poly(A)+RNA extracted from tilapia Oreochromis niloticus anterior pituitaries. The recombinant clones carrying the cDNA sequence of tilapia growth hormone (tiGH) were ... [more ▼]

A cDNA library was prepared from poly(A)+RNA extracted from tilapia Oreochromis niloticus anterior pituitaries. The recombinant clones carrying the cDNA sequence of tilapia growth hormone (tiGH) were selected using a fragment of the trout growth hormone (tGH) cDNA as hybridization probe. The nucleotide sequence of the full-length tiGH cDNA was determined. This cDNA encodes a protein of 204 amino acids, including the putative signal peptide of 17 amino acids. Mature tiGH cDNA was inserted in an Escherichia coli expression vector which led to the production of tiGH protein with a yield estimated to be 20% of the total bacterial proteins. [less ▲]

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See detailThe human genome contains hundreds of genes coding for finger proteins of the Kruppel type
Bellefroid, Eric J.; Lecocq, P.; Benhida, A. et al

in DNA (1989), 8(6), 377-87

Our aim was to identify new human proteins with potential DNA binding activity, related to the Kruppel protein which regulates Drosophila segmentation. We screened a human placenta cDNA library and a ... [more ▼]

Our aim was to identify new human proteins with potential DNA binding activity, related to the Kruppel protein which regulates Drosophila segmentation. We screened a human placenta cDNA library and a human genomic DNA library with a synthetic oligonucleotide probe corresponding to the H/C link region that connects finger loops in the multifingered Kruppel protein. We found more than 100 different mRNAs encoding Kruppel multifingered proteins in the human placenta. In the whole human genome, the number of genes encoding such proteins reaches about 300. Sequence analysis of 14 cloned cDNAs indicated that they code for at least nine undescribed human finger proteins. The sequences of the 106 finger repeats present in these nine proteins are highly homologous. Most of the variability lies in a limited number of positions located in their postulated alpha-helical structure, and therefore could be implicated in their DNA-binding specificity. [less ▲]

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See detailBinding of the human glucocorticoid receptor to defined regions in the human growth hormone and placental lactogen genes
Eliard, P. H.; Marchand, M.; Rousseau, G. G. et al

in DNA (1985), 4(6), 409-17

An in vitro competition assay was used to investigate whether binding sites for the human glucocorticoid receptor occur in the human genes for growth hormone (hGH) and placental lactogen (chorionic ... [more ▼]

An in vitro competition assay was used to investigate whether binding sites for the human glucocorticoid receptor occur in the human genes for growth hormone (hGH) and placental lactogen (chorionic somatomammotropin, hCS). These genes display 95% sequence homology. Two receptor-binding regions were found in the hGH gene, one of which is located within 290 bp upstream, and one within 251 bp downstream from the transcription initiation site. Two binding regions homologous to those in the hGH gene were found in the hCS gene. The receptor-binding DNA fragment from the structural part of the genes, but not that from their promoter area, contained a sequence homologous to a 15-bp consensus sequence proposed earlier for the glucocorticoid receptor binding site. It is unlikely that the putative difference in glucocorticoid sensitivity between the hGH and hCS genes is accounted for by major differences in glucocorticoid receptor binding pattern. [less ▲]

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See detailMolecular cloning of bovine viral diarrhea viral sequences
Renard, A.; Guiot, C.; Schmetz, D. et al

in DNA (1985), 4(6), 429-38

Bovine viral diarrhea virus (BVDV) genomic RNA was identified as a 12.5-kb single-stranded RNA molecule in both infected bovine embryonic kidney cells (BEK-1) and partially purified virions. BVD virion ... [more ▼]

Bovine viral diarrhea virus (BVDV) genomic RNA was identified as a 12.5-kb single-stranded RNA molecule in both infected bovine embryonic kidney cells (BEK-1) and partially purified virions. BVD virion RNA was partially purified and used as a template for cDNA synthesis. BVDV-specific cDNA sequences were molecularly cloned and shown to hybridize to infected cell RNA but not to uninfected cell RNA or DNA. A single RNA species of 12.5 kb, representing the viral RNA genome, was detected in infected cells. A preliminary map of the BVDV specific cDNA clones was constructed and five major, nonoverlapping families were observed, accounting for approximately one-half of the viral genome. [less ▲]

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See detailExpression and hormonal regulation of the rat growth hormone gene in transfected mouse L cells
Karin, M.; Eberhardt, N. L.; Mellon, S. H. et al

in DNA (1984), 3(2), 147-55

Expression of the rat growth hormone (rGH) gene in the pituitary and in cultured pituitary tumor cells is regulated by glucocorticoid hormones. After co-transfer of cloned DNA containing the rGH gene with ... [more ▼]

Expression of the rat growth hormone (rGH) gene in the pituitary and in cultured pituitary tumor cells is regulated by glucocorticoid hormones. After co-transfer of cloned DNA containing the rGH gene with the herpes simplex virus (HSV) thymidine kinase (tk) gene into mouse Ltk- cells, rGH gene transcripts were detected in eight of fifteen tk+ cell lines. However, in all eight clones, the predominant rGH gene transcript was only about 0.75 kb, 0.3 kb shorter than pituitary rGH mRNA. The 0.75-kb transcripts, examined from one clone, L-rGH-4, lacked sequences derived from exons 1 and 2 of the rGH gene. Although transcripts larger than 0.75 kb were detected, the normal 2.2-kb rGH gene primary transcript was present only at very low levels. Nuclease mapping studies also failed to reveal transcripts initiated at the normal rGH gene promoter, but instead revealed transcripts with 5' termini arising within intron B of the gene. These data suggest either that transcripts arise from internal promoters within the rGH gene or that a transcript initiated upstream from the normal promoter was processed abnormally. Dexamethasone increased the levels of the 0.75-kb rGH gene transcripts about fourfold in all eight clones expressing rGH mRNA. These data suggest that structural elements important for glucocorticoid-mediated influences on regulation of GH gene expression are contained within the transferred rGH gene fragment and can function even when the normal rGH gene promoter is not used and the pattern of expression is grossly abnormal. [less ▲]

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See detailA method for isolation of intact, translationally active ribonucleic acid
Cathala, G.; Savouret, J. F.; Mendez, B. et al

in DNA (1983), 2(4), 329-35

A method for isolation of large, translationally active RNA species is presented. The procedure involves homogenization of cells or tissues in 5 M guanidine monothiocyanate followed by direct ... [more ▼]

A method for isolation of large, translationally active RNA species is presented. The procedure involves homogenization of cells or tissues in 5 M guanidine monothiocyanate followed by direct precipitation of RNA from the guanidinium by 4 M LiCl. Modifications are described for use with tissue culture cells, yeast, tissues, or isolated nuclei. The advantages of the procedure include speed, simplicity, avoidance of an ultracentrifugation, and its applicability to large numbers of small samples. The procedure yields large mRNA precursors up to 10 kb and mRNA species which translate very well. However, small (less than 300 nucleotides) RNA species are recovered with a poor yield. [less ▲]

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See detailHormonal regulation of growth hormone mRNA
Wegnez, M.; Schachter, B. S.; Baxter, J. D. et al

in DNA (1982), 1(2), 145-53

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See detailCloning of bovine prolactin cDNA and evolutionary implications of its sequence
Miller, W. L.; Coit, D.; Baxter, J. D. et al

in DNA (1981), 1(1), 37-50

Prolactin, growth hormone, and chorionic somatomammotropin (placental lactogen) constitute a set of related polypeptides believed to derive from a common evolutionary ancestor protein. We have cloned and ... [more ▼]

Prolactin, growth hormone, and chorionic somatomammotropin (placental lactogen) constitute a set of related polypeptides believed to derive from a common evolutionary ancestor protein. We have cloned and sequenced DNA complementary to the mRNA coding for bovine prolactin. This cDNA contains 702 bases corresponding to 10 amino acids in the leader peptide, all 199 amino acids of the hormone, and 75 nucleotides in the 3' untranslated region of the mRNA. Nucleotide sequence analysis of this cDNA permitted the identification of 10 amino acids in the signal peptide, plus the correction or elucidation of amino acid assignments at 16 sites where aspartic and glutamic acids had not been distinguished from their amides by amino acid sequencing. Codon usage in bovine prolactin mRNA is nonrandom, but, similarly to rat and human prolactins, it does not exhibit the strong preference for G or C in codon third positions seen in bovine, rat, and human growth hormone mRNAs. The translational termination signal in bovine prolactin in UAA, also the same as in rat and human prolactins and differing from the UAG "stop" codon used in bovine, rat, and human growth hormones and human chorionic somatomammotropin. The amino acid and mRNA nucleotide sequences of bovine, rat, and human prolactins and growth hormones were compared by several techniques based on various theories of molecular evolution. The comparison of prolactin to growth hormone is consistent in all three species, suggesting that the genes for these two hormones diverged about 350 million years ago. However, comparisons among the three prolactins or among the three growth hormones to determine the times of evolutionary divergence of the three species generated values that were inconsistent with each other and with the fossil record. Analysis of these discrepancies suggests that the genes for prolactin and growth hormone may now be evolving by different mechanisms. [less ▲]

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