Effective transformation of the cyanobacterium Spirulina platensis using electroporation

Although Spirulina (Arthrospira) is expected to be a suitableorganism for producing recombinant proteins, a gene transfer system hasnot yet been established, due to a lack of suitable vectors and because Spirulina appears refractory to common genetic manipulations. As theinitial stages of the development of recombinant DNA methodology, weexamined the effects on transformation efficiency of electroporationconditions such as electric-field strength (2, 4, 6, 8, 10, 12kV cm^-1) and time constant (2.5, 5 ms). At a time constant of2.5 ms, few transformants were observed regardless of the field strength.The longer time constant of 5.0 ms reproducibly yielded transformants atthe middle field strength of 4 - 8 kV cm^-1, but gave high killingand no transformation at the higher field strength of 10 - 12kV cm^-1. Chloramphenicol acetyltransferase (CAT) activities wereincreased only in the transformants from 2–6 kV cm^-1 and 5.0 ms.The density of the transformants was significantly correlated with therelative value of CAT activity (r = 0.89, n = 11, p < 0.01),suggesting that the chloramphenicol resistance was due to CAT activity. Weconcluded that transformation of S. platensis was most effective at apulse duration 5.0 ms with an electric field of 4 kV cm^-1, and thatforeign genes can be expressed in this organism.