BioTechniques (Jan 2012)

Generating a custom TA-cloning expression plasmid for Lactococcus lactis

  • Aleš Berlec,
  • Borut Štrukelj

DOI
https://doi.org/10.2144/000113800
Journal volume & issue
Vol. 52, no. 1
pp. 51 – 53

Abstract

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The growing popularity of the lactic acid bacterium Lactococcus lactis has increased demand for novel high-throughput cloning methods. Here we describe a general TA-cloning methodology and demonstrate its feasibility using the plasmid pNZ8148. PCR products were directly ligated into a linear, PCR-amplified and XcmI-digested pNZ8148 derivative that was termed pNZ-T. Cloning using pNZ-T yielded a high proportion of insert-containing plasmids on transformation. Although demonstrated with L. lactis, the technique presented here is organism-independent and can be implemented in other plasmids.

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