ABSTRACT
A mutation in lac-operator region of pUC19 plasmid causing an increase in beta-galactosidase activity was observed. The plasmid was used as a vector to provide high level of expression of the cloned E. coli rplL gene.
Subject(s)
Escherichia coli/genetics , Gene Expression Regulation, Bacterial , Genes, Bacterial , Plasmids/genetics , Cloning, Molecular/methods , DNA, Bacterial/genetics , DNA, Bacterial/isolation & purification , Escherichia coli/enzymology , Genetic Vectors , Lac Operon , Mutation , Phenotype , beta-Galactosidase/geneticsABSTRACT
The possibility of the translation of mRNA isolated from bovine brain and injected into Xenopus oocytes has been shown. It is found that mRNA injection induced the appearance of voltage-operated TTX-sensitive sodium and calcium currents, GABA receptors in oocytes membrane.
Subject(s)
Brain/metabolism , Oocytes/drug effects , Poly A/pharmacology , RNA, Messenger/pharmacology , Xenopus/physiology , Animals , Calcium Channels/drug effects , Calcium Channels/physiology , Cattle , Female , Membrane Potentials/drug effects , Microbial Collagenase/pharmacology , Oocytes/physiology , Poly A/isolation & purification , Protein Biosynthesis/drug effects , RNA, Messenger/isolation & purification , Sodium Channels/drug effects , Sodium Channels/physiology , Tetrodotoxin/pharmacologyABSTRACT
An attempt has been undertaken to review the present data on the structure of alpha- and beta-like human globin gene families, their quantity in a cell genome, intergene spacers, evolution and common features and differences in globin genes sequences. Some other questions, concerning the regulation of globin genes expression, transcription, structure and processing of primary transcripts, the structure of globin messenger RNAs have been also discussed.
Subject(s)
Genes , Globins/genetics , Base Sequence , DNA/genetics , DNA Restriction Enzymes , Humans , RNA, Messenger/genetics , Transcription, GeneticABSTRACT
cDNA synthesized on rabbit bone marrow erythroid cells pre-mRNA was cloned in bacterial plasmids. Cold phenol extracted pre-mRNA was a several times more effective template in the reaction of reverse transcription without oligo(dT) 10-primer than hot phenol extracted pre-mRNA. There was no yield increase of DNA-product on hot phenol extracted pre-mRNA in the reaction of reverse transcription with the oligo(dT)10-primer addition. The "hot phenol" poly (A)+-pre-mRNA was used to obtain the representative, full-sized cDNA. The double-stranded form of this cDNA, obtained with the help of DNA-polymerase I, was inserted into the PstI-site of pBR322 plasmid. About 25% E. coli JC5183 clones, transformed with this hybrid plasmid, were found to contain the globin sequences.
