Integration of growth hormone gene constructs in transgenic strains of coho salmon (Oncorhynchus kisutch) at centromeric or telomeric sites

Genome ◽  
2010 ◽  
Vol 53 (1) ◽  
pp. 79-82 ◽  
Author(s):  
Ruth B. Phillips ◽  
Robert H. Devlin

Very little information is currently available regarding the sites of integration of transgenes in genetically engineered fish. We examined the chromosomal location of growth hormone gene constructs containing GH1 in three different strains of transgenic coho salmon produced by microinjection into pronuclei of fertilized eggs. The constructs were labeled and used as probes in fluorescence in situ hybridization experiments on chromosome preparations from the M77, MT5750A, and H3D0474 strains of transgenic coho salmon. The constructs were localized at 1–3 different sites in different strains. In the M77 strain the construct was found at a single centromeric site on a medium-sized metacentric chromosome, while in the MT5750A strain, the construct was found at a single telomeric site on the short arm of chromosome pair 21, a subtelocentric chromosome with a large band of repetitive DNA on the short arm. In the H3D0474 strain, the construct was found at telomeric sites on the long arms of three metacentric chromosomes that appear to represent one pair of homologous chromosomes and one chromosome containing the homeologous long arm (recently duplicated chromosome arm) corresponding to the long arm of the first pair. This suggests transfer of the construct may have occurred by homologous and homeologous crossing over. All of the constructs incorporated at restricted sites characterized by the presence of tandem DNA repeats.

2007 ◽  
Vol 50 (1) ◽  
pp. 141-146 ◽  
Author(s):  
Rafael Augusto de Carvalho ◽  
Ana Lúcia Dias

Twenty-five specimens of Iheringichthys labrosus from the Capivara Reservoir were analysed cytogenetically. AgNORs were detected in a pair of ST chromosomes, in the telomeric region of the long arm. Some individuals showed size heteromorphism of this region between homologous chromosomes. Treatment with CMA3 displayed GC-rich regions corresponding to the AgNOR pair, plus other fluorescent staining. In situ hybridization by fluorescence (FISH) with the 18S rDNA probe showed only one pair of stained chromosomes, confirming the heteromorphism observed with AgNO3 and CMA3 in some individuals. The 5S rDNA probe revealed telomeric staining on the long arm of a pair of chromosomes of the ST-A group, probably different from the NOR pair.


2000 ◽  
Vol 57 (5) ◽  
pp. 939-950 ◽  
Author(s):  
James A Hill ◽  
Anders Kiessling ◽  
Robert H Devlin

Transgenic coho salmon (Oncorhynchus kisutch) containing a growth hormone gene construct were compared with nontransgenic coho salmon in terms of gross anatomy, muscle cellularity, muscle enzyme activity, and differential gene expression. Transgenic fish were found to have significantly higher numbers of small-diameter muscle fibres in both the dorsal and lateral region of the somitic muscle, suggesting that they grow by greater rates of hyperplasia relative to slower growing nontransgenic fish. Higher levels of activity were found for phosphofructokinase and cytochrome oxidase in white muscle of the transgenic fish. This difference indicates a higher glycolytic and aerobic requirement in the muscle of transgenic fish. Subtractive hybridisation of muscle RNA of transgenic fish from control fish provided a library of cDNAs whose expression is upregulated in the transgenic fish. This library contains genes that may be involved in, or related to, both high growth rates and muscle hyperplasia. We have sequenced a number of fragments and have found a preponderance of myosin light chain 2 mRNAs, consistent with a putative high level of expression in the early stages of muscle fibre construction.


1991 ◽  
Vol 82 (6) ◽  
pp. 505-508 ◽  
Author(s):  
E.M. Shaw ◽  
R.N. Shoffner ◽  
D.N. Foster ◽  
K.S. Guise

1987 ◽  
Vol 75 (4) ◽  
pp. 326-332 ◽  
Author(s):  
Alessandra Agresti ◽  
G. Rainaldi ◽  
Andrea Lobbiani ◽  
Ivana Magnani ◽  
R. Di Lernia ◽  
...  

Genomics ◽  
1990 ◽  
Vol 8 (1) ◽  
pp. 171-174 ◽  
Author(s):  
Robert Hediger ◽  
Sam E. Johnson ◽  
William Barendse ◽  
Roger D. Drinkwater ◽  
Stephen S. Moore ◽  
...  

1998 ◽  
Vol 141 (1) ◽  
pp. 5-20 ◽  
Author(s):  
Jennifer C. Fung ◽  
Wallace F. Marshall ◽  
Abby Dernburg ◽  
David A. Agard ◽  
John W. Sedat

The dynamics by which homologous chromosomes pair is currently unknown. Here, we use fluorescence in situ hybridization in combination with three-dimensional optical microscopy to show that homologous pairing of the somatic chromosome arm 2L in Drosophila occurs by independent initiation of pairing at discrete loci rather than by a processive zippering of sites along the length of chromosome. By evaluating the pairing frequencies of 11 loci on chromosome arm 2L over several timepoints during Drosophila embryonic development, we show that all 11 loci are paired very early in Drosophila development, within 13 h after egg deposition. To elucidate whether such pairing occurs by directed or undirected motion, we analyzed the pairing kinetics of histone loci during nuclear cycle 14. By measuring changes of nuclear length and correlating these changes with progression of time during cycle 14, we were able to express the pairing frequency and distance between homologous loci as a function of time. Comparing the experimentally determined dynamics of pairing to simulations based on previously proposed models of pairing motion, we show that the observed pairing kinetics are most consistent with a constrained random walk model and not consistent with a directed motion model. Thus, we conclude that simple random contacts through diffusion could suffice to allow pairing of homologous sites.


1988 ◽  
Vol 263 (11) ◽  
pp. 5005-5007 ◽  
Author(s):  
L N Peritz ◽  
E J Fodor ◽  
D W Silversides ◽  
P A Cattini ◽  
J D Baxter ◽  
...  

Genes ◽  
2021 ◽  
Vol 12 (8) ◽  
pp. 1128
Author(s):  
Marilena Nakaguma ◽  
Nathalia Garcia Bianchi Pereira Ferreira ◽  
Anna Flavia Figueredo Benedetti ◽  
Mariana Cotarelli Madi ◽  
Juliana Moreira Silva ◽  
...  

We report four allelic variants (three novel) in three genes previously established as causal for hypopituitarism or related disorders. A novel homozygous variant in the growth hormone gene, GH1 c.171delT (p.Phe 57Leufs*43), was found in a male patient with severe isolated growth hormone deficiency (IGHD) born to consanguineous parents. A hemizygous SOX3 allelic variant (p.Met304Ile) was found in a male patient with IGHD and hypoplastic anterior pituitary. YASARA, a tool to evaluate protein stability, suggests that p.Met304Ile destabilizes the SOX3 protein (ΔΔG = 2.49 kcal/mol). A rare, heterozygous missense variant in the TALE homeobox protein gene, TGIF1 (c.268C>T:p.Arg90Cys) was found in a patient with combined pituitary hormone deficiency (CPHD), diabetes insipidus, and syndromic features of holoprosencephaly (HPE). This variant was previously reported in a patient with severe holoprosencephaly and shown to affect TGIF1 function. A novel heterozygous TGIF1 variant (c.82T>C:p.Ser28Pro) was identified in a patient with CPHD, pituitary aplasia and ectopic posterior lobe. Both TGIF1 variants have an autosomal dominant pattern of inheritance with incomplete penetrance. In conclusion, we have found allelic variants in three genes in hypopituitarism patients. We discuss these variants and associated patient phenotypes in relation to previously reported variants in these genes, expanding our knowledge of the phenotypic spectrum in patient populations.


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