[Cloning and expression of the B1 hordein gene of barley (Hordeum vulgare L.) in cells of the cyanobacterium Synechocystis sp. PCC6803 and Escherichia coli].

The hordein B1 gene of Hordeum vulgare L. (variety Donetskii 4) was cloned 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 carrying the hordein B1 gene was inserted into plasmid pBSM13(-), controlled by a lac promoter. The constructed recombinant plasmid pBSB1 provides hordein B1 gene expression in transformed E. coli cells. To introduce the hordein B1 gene into the genome of Synechocystis sp. PCC6803, an integrative vector, containing a fragment of cyanobacterium chromosomal DNA with inserted Tn5 antibiotic resistance genes (KmR, BleoR, StpR) was constructed. A Pvu II-fragment of pBSB1, containing hordein B1 gene under control of the lac promoter, was cloned in the Sma I-site of gene BleoR. The resulting recombinant plasmid was used to transform cyanobacterial cells. One KmR transformants contained a DNA fragment yielding a positive signal during Southern blotting with a [32P]-labeled DNA fragment carrying hordein B1 gene. Western blotting with polyclonal antibodies to barley hordein revealed hordein B1 gene expression in Synechocystis sp. PCC6803.