ABSTRACT
Effect of complementary oligonucleotides and their reactive derivatives on translation of mouse immunoglobulin G kappa light chain was investigated. It was found that oligonucleotide pTGCTCTGGTTT and shorter oligonucleotides complementary to the coding sequence of the mRNA (nucleotides 205-215) do not arrest translation of the mRNA in the rabbit reticulocyte cell-free translation system. Preincubation of the mRNA with the alkylating 4-(N-2-chloroethyl-N-methylamino)benzyl-5'-phosphamide derivative of the oligonucleotide completely suppresses the synthesis of the protein thus demonstrating higher efficiency of the reactive oligonucleotide derivatives as inhibitors of the mRNA function.
Subject(s)
Alkylating Agents/pharmacology , Immunoglobulin G/genetics , Oligonucleotides/pharmacology , Protein Biosynthesis/drug effects , Protein Synthesis Inhibitors , RNA, Messenger/genetics , Animals , Cell-Free System , Electrophoresis, Polyacrylamide Gel , Immunoglobulin G/biosynthesis , Immunoglobulin Heavy Chains/biosynthesis , Immunoglobulin Heavy Chains/genetics , Immunoglobulin Light Chains/biosynthesis , Immunoglobulin Light Chains/genetics , Mice , Rabbits , Reticulocytes/metabolismABSTRACT
Characterization of 12 clones of monoclonal antibodies (MAb) generated for the main immunogen of tick-borne encephalitis virus, glycoprotein E, is presented. The following MAb parameters have been determined: constants of binding with antigen, classes and subclasses of immunoglobulins, the activity in two variants of solid-phase enzyme-immunoassay, binding with protein A, and MAb behavior in serologic tests: hemagglutination-inhibition, diffuse precipitation in agar, and virus neutralization. The preliminary studies revealed the presence in these MAb of at least three groups of antibody complementary to various nonoverlapping sites on the structural virus glycoprotein. Further employment of these MAb in practical and research work is discussed.
Subject(s)
Antibodies, Monoclonal/isolation & purification , Encephalitis Viruses, Tick-Borne/immunology , Glycoproteins/immunology , Viral Structural Proteins/immunology , Animals , Enzyme-Linked Immunosorbent Assay , Hybridomas , Immunodiffusion , Mice , Mice, Inbred BALB CABSTRACT
Sequence specific modification of nucleic acids with reactive oligonucleotide derivatives, complementary addressed modification, can provide an efficient approach for specific inactivation of certain cellular nucleic acids. In experiments with ascites tumor Krebs II cells and alkylating oligothymidylate derivatives it was found that alkylating oligonucleotide derivatives enter the living cell and modify complementary sequences in cellular nucleic acids with high efficiency. Complementary addressed modification of poly(A) sequences in cellular RNA with oligothymidylate derivatives was investigated in detail. The results of experiments on alkylation of cellular nucleic acids are consistent with complementary addressed modification of poly(A) sequences in cellular DNA. These results are supported by experiments on modification of chromatin DNA in which it was found that chromatin DNA interacts with oliogothymidylate derivatives more readily than the isolated double stranded DNA. It was found that alkylating oligonucleotide derivatives complementary to a sequence in immunoglobulin mRNA of MOPC 21 cells arrest the cellular immunoglobulin synthesis. Alkylating oligonucleotide derivatives complementary to RNAs of fowl plague virus inhibit virus multiplication in cell culture.
Subject(s)
Nucleic Acids/antagonists & inhibitors , Oligonucleotides/pharmacology , Alkylation , Animals , Base Sequence/drug effects , Carcinoma, Krebs 2/metabolism , Chromatin/drug effects , Chromatin/metabolism , Immunoglobulin G/metabolism , Influenza A virus/drug effects , Mice , Mice, Inbred Strains , Multiple Myeloma/immunology , RNA, Messenger/immunologySubject(s)
Alkylating Agents/pharmacology , Genetic Code/drug effects , Immunoglobulin Light Chains/biosynthesis , Oligonucleotides/pharmacology , Plasmacytoma/immunology , RNA, Messenger/genetics , RNA, Neoplasm/genetics , Animals , Cells, Cultured , Depression, Chemical , Immunoglobulin Light Chains/genetics , Mice , Mice, Inbred BALB CABSTRACT
Polynucleotides adsorb on natural and model phospholipid membranes in the presence of Mg2+-cations. Adsorption of nucleic acids on membranes results in a considerable change of their secondary structure. The presence of model phosphatidylcholine membranes greatly stimulates the rate of the synthesis of RNA by E. coli RNA-polymerase on DNA template.
