Integration of Emphasis Type="Italic"hup/Emphas

Authors: N Vijaya Bhanu S P S Khanuja M L Lodha

Publish Date: 2013/04/15

Volume: 3, Issue: 1, Pages: 19-24

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Abstract

The hup gene fragment of cosmid pHU52 was integrated into the genome of chickpeaRhizobium Rcd301 via sitespecific homologous recombination Two small fragments of genomic DNA of strain Rcd301 itself were provided to flank cloned hup genes to facilitate the integration The hup insert DNA of cosmid pHU52 was Isolated as an Intact 302 kb fragment using EcoRI and cloned on partially restricted cosmid clone pSPSm3 which carries a DNA fragment of strain Rcd301 imparting streptomycin resistance One of the recombinant cosmid clones pBSL 12 thus obtained was conjugally transferred to the strain Rcd301 The integration of hup gene fragment into the genomic DNA through sitespecific homologous recombination was ensured by introducing an incompatible plasmid pPH1 JI The integration was confirmed by Southern hybridization The integrated hup genes were found to express ex plants in two such constructs BSL 12–1 and BSL 12–3

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