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Development of genetic markers in cyanobacteria and stability of genetically marked strains in soil
Journal article   Peer reviewed

Development of genetic markers in cyanobacteria and stability of genetically marked strains in soil

Sankar Subramanian and B D Kaushik
World Journal of Microbiology and Biotechnology, Vol.17(5), pp.535-544
2001
DOI: https://doi.org/10.1023/A:1011940212656
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https://doi.org/10.1023/A:1011940212656View
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Abstract

Biological Sciences Technology cyanobacteria genetic marker GUS luc marker marked strain's stability triparental conjugation vector
Reporter marker GUS (β-glucuronidase gene from Escherichia coli) and luc operon from the American firefly were introduced into cyanobacteria and the stability of these markers in soil was examined. To transfer the integrational vector into cyanobacteria, the genomic DNA library of Synechocystis sp. or Anabaena cylindrica maintained in pBR 322, pCY 100 and pCY 101 were transformed with HB 101 containing pRL 528 and selected for Cmr and Ampr. These clones of HB 101 containing pRL 528 and the vectors carrying different cyanobacterial chromosomal DNA fragments were used for triparental mating with HB 101 [pRK 2013/pRK 2073] and cyanobacteria. The frequency of transconjugants for integrational vectors was between 2.1 × 10-5 and 4.0 × 10-4. The transfer frequency of RSF 1010 based vectors (pDSK 519 and pCY 106) was 1.0-4.5 × 10-4 in Synechocystis sp. whereas A. cylindrica failed to maintain these vectors. Low frequency transfer (2.0-2.3 × 10-6) of RK 2 based vectors pVK 100 and pCY 104 was observed in A. cylindrica but these were unable to replicate in Synechocystis sp. The vectors in general were stable at least by 74.9% for 60 days of incubation in BG-11 medium. The markers were less stable in A. cylindrica (74.9-84.2%) compared to Synechocystis sp. (80.1-88.8%) at 60 days of incubation. Integrational vectors were almost 85% stable in both the strains. The RK 2 derivative of pCY 104 was less stable in A. cylindrica (74.9-77.3%) than the RSF 1010-based vector pCY 106 in Synechocystis sp. (80.1-81.0%). A maximum of 64.7% of the markers were lost in soil. The chromosomal markers through integrational vectors were found to be highly stable and 68.2-72.7% of these markers were retained in cyanobacteria at 60 days of incubation. Plasmid markers were less stable, with a loss of 64.7-48.7% at the end of the experiment. In A. cylindrica 58-65% of the RK 2 vector was lost whereas in Synechocystis sp. 49-61% of RSF 1010 was lost at 60 days of incubation.

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