[Interferon secreted by Bacillus subtilis is retained by membranes].
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Biomedical subjects
Publications and source records attributed to E G Abalakina.
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The regions of B6K DNA corresponding to oriT, finO and traS are mapped using a number of hybrid plasmids and deletion mutants pAS3::Tn5, pAS3::Tn7 and pAS3::Tn9 obtained in vitro after treatment with restriction endonucleases EcoRI and BamHI. FinO- and traS-like genes were mapped in the regions of 10 and 25,0-25,4 MD, respectively, oriT being situated in the region of 4,6-4,7 Md.
The regions determining conjugation ability of plasmid R6K were localized by means of deletion mutants obtained in vitro and in vivo and Tra- mutants induced by integration of transposons Tn5, Tn7 and Tn9 into different DNA sites of the conjugative deletion mutant pAS3. At least 13 genes were found to be involved in the genetic control of R6K conjugativity, on the basis of genetic, restriction and heteroduplex studies. They were mapped within the two DNA regions having the total molecular weight of about 10-11 md. A transposon-like structure with a replication function has been located between them.
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The results of genetic studies of R6K Tra1- and R6Kdelta[Sm1] mutants of R6K plasmid and those of heteroduplex analysis of DNAs have shown that DNA of this drug-resistant factor contains three loops flanked by the inverted repeats. The latter are designated as IR1, IR2 and IR3 and are of 50, 100 and 120 nucleotides in size respectively. IR1 is inserted into the loop flanked by IR2. Loops with these two repeats are located in major EcoR1 fragment, IR3 having been found in minor EcoRI fragment of the plasmid. The evidence obtained from the analysis of heteroduplex R6K/RSF2124 has shown that the loop with IR1 is corresponding to transposon Tn3. The extent of the deletion deltaSm1 indicates that IR2 may be a part of a transposon bearing the resistance to streptomycin. By comparing present data with those obtaine from the analysis of the RSF1040 factor of DNA replication initiation sites (Grosa et al., 1976), it has been suggested that the loop with IR3 represents a transposon with replicative functions (TnRep). The deletion of the mutant plasmid R6Kdelta[Sm1] (7.2 . 10(6) daltons in size) which affected one of the EcoRI sites not only confers the sensitivity to streptomycin but enhances also the efficiency of conjugational transfer and results in the loss of the R6K ability to bring about integrative suppression and to inhibit the fertility of the plasmids from IncP and IncN groups. The deletion mutant proved to have lost the property of incompatibility with the initial plasmid R6K and with itself.
Alpha-amylase genes of Bacillus amyloliquefaciens, coding proteins with reduced thermostability, had been obtained as a result of hydroxylamine mutagenesis. Temperature, pH and starch concentration dependences of two mutant alpha-amylases were investigated. The synthesis of the alpha-amylases by several B. subtilis strains with different levels of extracellular proteases was also studied. The mutation containing fragments were localized and the structures of the mutations were determined. It was found that the decrease of thermostability of mutant No 141 was due to Asp to Asn change at the position No 194 of the mature protein, and for mutant No 191--due to Glu to Lys change at the position No 185.
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