CLONING AND EXPRESSION OF HORDEIN B1-GENE OF BARLEY (HORDEUM-VULGARE L) IN CYANOBACTERIUM-SYNECHOCYSTIS SP-PCC6803 AND ESCHERICHIA-COLI

The hordein B1 gene of Hordeum vulgare L. (variety Donetskii 4) was cl oned in cells of Escherichia coli and cyanobacterium Synechocystis sp. PCC6803. For cloning in E. coli, a 2.3-kb Hpa I/EcoR I DNA fragment c arrying the hordein B1 gene was inserted into plasmid pBSM13(-), contr olled by a lac promoter. The constructed recombinant plasmid pBSB1 pro vides hordein B1 gene expression in transformed E. coli cells. To intr oduce the hordein B1 gene into the genome of Synechocystis sp. PCC6803 , an integrative vector, containing a fragment of cyanobacterium chrom osomal DNA with inserted Tn5 antibiotic resistance genes (Km(R), Bleo( R), Stp(R)) was constructed. A Pvu II-fragment of pBSB1, containing ho rdein B1 gene under control of the lac promoter, was cloned in the Sma I-site of gene Bleo(R). The resulting recombinant plasmid was used to transform cyanobacterial cells. One Km(R) transformants contained a D NA fragment yielding a positive signal during Southern blotting with a [P-32]-labeled DNA fragment carrying hordein B1 gene. Western blottin g with polyclonal antibodies to barley hordein revealed hordein B1 gen e expression in Synechocystis sp. PCC6803.