This paper represent the second step i n a molecular clon i ng program ai ming to clone large DNA fi·agmen ts of the sal t tolerant bermudagrass (Cyrwdon dactylon L.) DNA usi ng the bacteriophage (EM13L3) as a vector.
In th is work, a yield of about I 00 g bacteriophage DNA per one liter culture.was obtained with.a purity ranging between (1.7-1.8). The vector JJNA v.as completely double digested with the restriction enzymes llamHI and EcoRI, followed by purification of vector arms. llacteriophage anns were also efficiently ligated with the partially digested bermudagrass DNA (prepared earlier). The optimum ligation
ratio or arms: inserts (EMBL3: bermudagrass) was found to be 4: I
respectively. The recombinant DNA was successfu lly in vitro packaged and plated on a P2 lysogen E. coli (strain N M539). An efficiency of about 1.3x I 06 pfu/ tg recombi l1311l ph age DNA was determined in these ex periments.