An investigation on gene transfer to spirulina platensis (Cyanophyta)

In this study, serin esterase gene from Spirulina platensis amplified by PCR using twodifferent primer sequences. The amplified gene cloned into the pBR325 vector in twodifferent way, depending on differences between primers, thus, two entegrative vectors for S.platensis were constructed. The new entegrative vectors were identified as pBR325SE1 andpBR325SE2 respectively. While pBR325SE1 and pBR325SE2 were transferred intocompetent E. coli, besides this, pBR325SE1 was electroporated into S. platensis.Recombinant S. platensis/pBR325SE1 were grown in the Spirulina medium withchloramphenicol (cml) while there wasn?t any non-transformant Spirulina in the samemedium but intensive non-transformant growth was observed in the medium without cml.Bacteriophage T4 lysosyme gene was cloned into pBR325SE2 and the new vectorharbouring lysozyme gene was identified as pBR325SL. pBR325SL was transferred intocompetent E. coli and S. platensis respectively. Recombinant S. platensis/pBR325SLobserved in the Spirulina medium with cml while there was no non-transformant Spirulina inthe same medium but intensive non-transformant growth was observed in the mediumwithout cml. While the presence of lysozyme gene was demonstrated in pBR325SL isolatedfrom E. coli by PCR, but it couldn?t be demonstrated in S. platensis/pBR325SL because ofinsufficient Spirulina in the medium with cml growth for DNA isolation.Key Words: Spirulina platensis, Serin esterase, Integrative vector, PCR, Cloning