Recombinant plasmids containing Xenopus laevis globin structural genes derived from complementary DNA

Humphries, Peter; Old, Robert; Coggins, Lesley W.; McShane, Theresa; Watson, Christine J.; Paul, John

doi:10.1093/nar/5.3.905

Cited by 47 publications

(11 citation statements)

References 27 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Blackler. Preparation and Purification of DNA High molecular weight genomic DNA was prepared from blood by RNAse and proteinase digestion of red cell nuclei, followed by phenol extraction (19). Supercoiled plasmid DNAs were isolated by the method of Colman et al (20) and inserts prepared by elution from agarose gel slices followed by DE52 chromatography.…”

Section: Materials and Methods Animalsmentioning

confidence: 99%

Histone gene number and organisation inXenopus: Xenopus borealishas a homogeneous major cluster

Turner

Woodland

1983

Nucl Acids Res

View full text Add to dashboard Cite

Section: Materials and Methods Animalsmentioning

confidence: 99%

Histone gene number and organisation inXenopus: Xenopus borealishas a homogeneous major cluster

Turner

Woodland

1983

Nucl Acids Res

View full text Add to dashboard Cite

“…Transfer of plaques or bacterial colonies to nitrocellulose filters (Schleicher and Schull) was done using the methods of Benton and Davis (1977). Hybridisation of 32P end-labelled RNA to the nitrocellulose bound DNA and subsequent washing were carried out by the methods of Humphries et al (1978). Restriction digests, ligations Restriction enzymes and T4 DNA ligase were obtained from New England Biolabs and used following the procedures of Maniatis et al (1982).…”

Section: Polyadenylation Of U Snrnasmentioning

confidence: 99%

Xenopus laevis U2 snRNA genes: tandemly repeated transcription units sharing 5′ and 3′ flanking homology with other RNA polymerase II transcribed genes.

Mattaj¹,

Zeller²

1983

The EMBO Journal

132

View full text Add to dashboard Cite

Xenopus laevis U2 small nuclear RNA (snRNA) genes were isolated and expressed by microinjection into frog oocytes. The genes are organised in short tandemly repeated units of approximately 830 bp. Some of the cloned tandem repeats are closely linked to genes coding for U5 snRNA, tRNA and an uncharacterised 7S RNA. No evidence was found for U2 snRNA pseudogenes. Single repeat units are transcriptionally active, showing that all the signals necessary for U2 snRNA transcription are included in an 831‐bp segment of DNA. Sequence analysis of a cloned repeat unit showed that Xenopus and rat U2 snRNAs are 94% homologous. Flanking regions 5′ and 3′ to the coding sequence were found which shared extensive homology with similarly positioned sequences in human U1 snRNA genes. Part of the 3′ non‐coding region homology (consensus TTTNAAAGAAT) was found in many other genes transcribed by RNA polymerase II.

show abstract

“…Restriction (4). Double-stranded cDNA with poly(dC) homopolymeric extensions was synthesized as described (9)(10)(11)(12).…”

mentioning

confidence: 99%

Nucleotide sequence of the mRNA encoding the pre-alpha-subunit of mouse thyrotropin.

Chin¹,

Kronenberg²,

Dee³

et al. 1981

Proc. Natl. Acad. Sci. U.S.A.

View full text Add to dashboard Cite

We have constructed and cloned in bacteria recombinant DNA molecules containing DNA sequences coding for the precursor of the a subunit of thyrotropin (pre-TSH-a). Double-stranded DNA complementary to total poly(A)+RNA derived from a mouse pituitary thyrotropic tumor was prepared enzymatically, inserted into the Pst I site of the plasmid pBR322 by using poly(dC)-poly(dG) homopolymeric extensions, and cloned in Escherichia coli x1776. Cloned cDNAs encoding pre-TSH-a were identified by their hybridization to pre-TSH-a mRNA as determined by cell-free translations of hybrid-selected and hybrid-arrested RNA. The nucleotide sequences oftwo cDNAs (510 and 480 base pairs) were determined with chemical methods and corresponded to much of the region coding for the a subunit and the 3' untranslated region of pre-TSH-a mRNA. The sequence of the 5' end of the mRNA was determined from cDNA synthesized by using total mRNA as template and a restriction enzyme DNA fragment as primer. Together these sequences represented >90% of the coding and noncoding regions of full-length pre-TSH-a mRNA, which was determined to be 800 bases long. The amino acid sequence of the pre-TSH-a deduced from the nucleotide sequence showed a NH2-terminal leader sequence of24 amino acids followed by the 96-amino-acid sequence ofthe apoprotein of TSHa. There is greater than 90% homology in the amino acid sequences among the murine, ruminant, and porcine a subunits and 75-80% homology among the murine, equine, and human a subunits. Several regions of the sequence remain absolutely conserved among all species, suggesting that these particular regions are essential for the biological function of the subunit. The successful cloning of the a subunit of TSH will permit further studies of the organization of the genes coding for the glycoprotein hormone subunits and the regulation of their expression.

show abstract

Recombinant plasmids containing Xenopus laevis globin structural genes derived from complementary DNA

Cited by 47 publications

References 27 publications

Histone gene number and organisation inXenopus: Xenopus borealishas a homogeneous major cluster

Histone gene number and organisation inXenopus: Xenopus borealishas a homogeneous major cluster

Xenopus laevis U2 snRNA genes: tandemly repeated transcription units sharing 5′ and 3′ flanking homology with other RNA polymerase II transcribed genes.

Nucleotide sequence of the mRNA encoding the pre-alpha-subunit of mouse thyrotropin.

Contact Info

Product

Resources

About