Construction and cloning of rat albumin structural gene sequences.
Open Access
- 1 September 1979
- journal article
- research article
- Published by Proceedings of the National Academy of Sciences in Proceedings of the National Academy of Sciences
- Vol. 76 (9), 4370-4374
- https://doi.org/10.1073/pnas.76.9.4370
Abstract
A recombinant plasmid containing a DNA segment complementary to rat liver albumin mRNA was constructed, cloned and used to examine the organization of albumin gene. The 18S fraction of total liver poly(A)-containing RNA was copied into a double-stranded c[complementary]DNA by avian myeloblastosis virus reverse transcriptase and Escherichia coli DNA polymerase I. The cDNA was inserted into the HindIII site of the plasmid pBR322 via the addition of specific oligonucleotide linkers. Recombinant plasmids were screened by hybrid arrest of mRNA translation and hybridization with specific cDNA. A plasmid was identified that contained a 1200-nucleotide insert corresponding to a segment adjacent to the 5''-terminal region of albumin mRNA. The inserted sequence was used as a hybridization probe to detect 5 EcoRI fragments of genomic DNA which encode albumin mRNA. These were compared to 8 EcoRI fragments identified within the rat genome by albumin cDNA. The albumin gene (or genes) is apparently interrupted at > 1 site in the coding DNA by intervening sequences. Those fragments that encode the 5'' and 3'' ends of the mRNA were distinguished.Keywords
This publication has 22 references indexed in Scilit:
- High capacity gel preparative electrophoresis for purification of fragments of genomic DNAAnalytical Biochemistry, 1978
- Purification and cloning of a mouse ribosomal gene fragment in coliphage lambdaGene, 1977
- Synthesis of a complementary DNA to rat liver albumin mRNABiochemical and Biophysical Research Communications, 1977
- Rat Insulin Genes: Construction of Plasmids Containing the Coding SequencesScience, 1977
- Protein expression in E. coli minicells by recombinant plasmidsCell, 1977
- Detection of specific sequences among DNA fragments separated by gel electrophoresisJournal of Molecular Biology, 1975
- Improved computer program data for the resolution and fractionation of macromolecules by isokinetic sucrose density gradient sedimentationAnalytical Biochemistry, 1974
- [68] The wheat embryo cell-free systemMethods in enzymology, 1974
- Cleavage of Structural Proteins during the Assembly of the Head of Bacteriophage T4Nature, 1970
- Sedimentation studies of the size and shape of DNAJournal of Molecular Biology, 1965