02067nas a2200229   4500008004100000022001400041245009900055210006900154260005700223300001400280490000700294520120800301653003101509653003001540653001701570653002501587100002401612700002401636700002701660700002201687856012801709       2008                        eng d  a0959-399300aIsolation and heterologous expression of PHA synthesising genes from Bacillus thuringiensis R10 aIsolation and heterologous expression of PHA synthesising genes   a233 SPRING ST, NEW YORK, NY 10013 USAbSPRINGERcSEP  a1769-17740 v243 a<p>The polyhydroxyalkanoate biosynthesis gene locus from Bacillus thuringiensis R1 was isolated, cloned and analyzed at the molecular level. We found that a similar to 5 kb SacI-ClaI digested fragment of genomic DNA from B. thuringiensis R1 encoding the PHA synthesising genes, conferred PHA producing ability to E. coli. The fragment was sequenced and found to be of 4787 bp with five open reading frames. Sequence alignment with closely related species of Bacillus in the existing database revealed that the ORFs correspond to phaP, phaQ, phaR, phaB and phaC genes. However, E. coli harboring phaP, phaQ, phaR, phaB and phaC locus produced very low PHA. Furthermore, complementation of the locus with phaA from Ralstonia eutropha increased the PHA production in the recombinant E. coli from 3.0% to 24% of cell dry mass. The putative promoter regions and ribosome binding sites were identified for each of the gene. Conserved domains for PHA synthase and aceto-acetyl-coA reductase were also identified. We hence conclude that the PHA operon of Bacillus thuringiensis R1 consists of phaP, phaQ, phaR, phaB, phaC and complementation of the same with phaA is accountable for its high PHA production.</p>10aaceto-acetyl-coA reductase10aBacillus thuringiensis R110aPHA synthase10apolyhydroxyalkanoate1 aDesetty, Rohini, D.1 aMahajan, Vineet, S.1 aKhan, Bashir, Mohammad1 aRawal, Shuban, K.  uhttps://library.ncl.res.in/content/isolation-and-heterologous-expression-pha-synthesising-genes-bacillus-thuringiensis-r1-0