Molecular cloning and in vitro transcription of rat 4.5S RNAh genes

Fumio Harada, Yasuhiro Takeuchi, Nobuyuki Kato

Research output: Contribution to journalArticle

10 Citations (Scopus)

Abstract

4.5S RNAH (4.5S RNA associated with poly A containing RNA) has extensive homology to major interspersed repeat Bl in rodent genomes. We developed a new cloning technique for screening genomic library that eliminates the signal produced by repeated sequences or pseudogenes and applied it to cloning of 4.5S RNAH genes. Six phage clones (2, 3, 6, 9, 10 and 15) which hybridize with 4.5S RNAH were isolated from a rat gene library by this method. The restriction fragments containing the 4.5S RNAH locus were subcloned into plasmids and sequenced. Clones 2, 3, 9 and 15 contained one to five base substitutions in the coding region for 4.5S RNAH and were probably pseudogenes. In clone 2, the 4.5S RNAH locus was linked directly with the identifier sequence. Clone 6 contained three copies of the 4.5S RNAH gene (6a, b and c) which were clustered in the same direction within 455 base pairs. 6b was linked directly with 6c and ubiquitous repetitive DNA sequences B2 were inserted immediately after 6a and 6c. These three sequences as well as the sequence in clone 10 were colinear with rat 4.5S RNAH. In an in vitro transcription system, only clone 10 gave intact 4.5S RNAH.

Original languageEnglish
Pages (from-to)1629-1642
Number of pages14
JournalNucleic acids research.
Volume14
Issue number4
DOIs
Publication statusPublished - Feb 25 1986
Externally publishedYes

Fingerprint

Cloning
Molecular Cloning
Transcription
Clone
Rats
Clone Cells
Genes
Gene
RNA
Pseudogenes
Poly A
Bacteriophages
Genomic Library
DNA sequences
Gene Library
Locus
Organism Cloning
Screening
Plasmids
Substitution reactions

ASJC Scopus subject areas

  • Statistics, Probability and Uncertainty
  • Applied Mathematics
  • Health, Toxicology and Mutagenesis
  • Toxicology
  • Genetics(clinical)
  • Genetics

Cite this

Molecular cloning and in vitro transcription of rat 4.5S RNAh genes. / Harada, Fumio; Takeuchi, Yasuhiro; Kato, Nobuyuki.

In: Nucleic acids research., Vol. 14, No. 4, 25.02.1986, p. 1629-1642.

Research output: Contribution to journalArticle

Harada, Fumio ; Takeuchi, Yasuhiro ; Kato, Nobuyuki. / Molecular cloning and in vitro transcription of rat 4.5S RNAh genes. In: Nucleic acids research. 1986 ; Vol. 14, No. 4. pp. 1629-1642.
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