Gene/Protein Disease Symptom Drug Enzyme Compound
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Query: EC:3.2.1.23 (beta-galactosidase)
 14,648 document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)

15 oligodeoxynucleotides were synthesized using the phosphotriether technique which were subsequently enzymatically ligated in polylinker with subsequent phasing of left and right sides. Based on the phased polylinker a series of vehicles for the gene cloning and expression was constructed. The vectors of pRK series contain all three variants of polylinker with the frame shift of the reading frame for 1, 2, or 3 nucleotides in both chains. The obtained polylinkers do not effect the enzymatic activity of the beta-galactosidase alpha-peptide. Structure of the phased polylinker was confirmed by Maxam and Gilbert's sequencing method.
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PMID:[A phased poly-linker for cloning and expression of genes]. 261 74

17- and 20-mer oligodeoxyribonucleotides and their analogues, containing one to four phosphate groups esterified with ethyl alcohol in different positions of oligonucleotide chain, were synthesized by modified triester method. Ethylated di- and trinucleotide blocks were prepared by transesterification method from chlorophenyl derivatives. The structures of the oligonucleotides were confirmed by Maxam-Gilbert sequencing method. Oligonucleotides were not totally complementary to the N-terminal region of lac Z'gene (coding for N-terminal fragment of beta-galactosidase) of phage M13mpB DNA and induced the formation of the proposed deletion mutant DNA M13mp1 delta T. Phosphotriester analogues were more effective mutagens as compared to phosphodiester oligonucleotides due to their stability to nucleases. The use of E. coli DNA-polymerase I provided the increase in the mutant yields in case of the phosphotriester analogues. The stability of the analogues to 5'----3'----5'-endonuclease action, the specificity of oligonucleotide: DNA binding and the structure of mutant DNA were studied by the Sanger sequencing method.
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PMID:[Site-localized mutagenesis directed by phosphotriester analogs of oligonucleotides]. 377 34

A recombinant plasmid pN62 determining the synthesis of a hybrid protein consisting of a full-size beta-galactosidase and C-terminus fragment of influenza A virus nucleoprotein was constructed. The complete identity of pN62 insert with the 3'-terminus cDNA fragment of influenza A virus NP-gene and conservation of beta-galactosidase reading frame was confirmed by Maxam-Gilbert sequencing of pN62. An expression of pN62 plasmid in E. coli JM103 in the presence of IPTG resulted in accumulation of fused protein as poorly soluble inclusion bodies in the bacterial cells. Analysis of E. coli JM103/pN62 bacteria lysates by 7% PAGE revealed that molecular weight of hybrid polypeptide was 18 kDa heavier than normal beta-galactosidase and corresponded to the previously deduced weight of 135 kDa.
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PMID:[The isolation and expression in E. coli of a recombinant plasmid containing the 3'-terminal fragment of the cDNA of the influenza A virus nucleoprotein gene]. 816 Apr 47

Analysis of published reports helped us single out the most potent antigens among HCMV proteins: phosphoproteins pp150(UL32) and p52(UL44). Theoretical computer analysis of p52 epitopes showed the main antigenic determinants not cross-reacting with antigens of other viruses. Virus-containing (strain AD169) material was obtained and genome DNA was isolated. Amplification of a site of gene UL44 coding for unique determinants detected a PCR fragment of required electrophoretic mobility. The fragment was cloned in vector pLBE. The specificity of cloning was confirmed by restriction analysis of theoretical sites. Nucleotide sequence of cloned fragment of UL44 gene was studied by Maxam-Gilbert's method. Cloning in expressing bacterial vectors helped obtain HCMV recombinant protein p52 in the pure form and fused with beta-galactosidase. Enzyme immunoassay with HCMV-positive and negative donor sera and ABBOTT HCMV sera showed that recombinant p52 increased the sensitivity and specificity of a previously obtained recombinant pp150 as an antigen to HCMV-IgG and HCMV-IgM. The sensitivity and specificity is 100% with 98-99% reliability.
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PMID:[Preparation of P52 recombinant antigenic protein from human cytomegalovirus (HCMV)]. 1118 55