Molecular cloning of large DNA fragments of the salt tolerant wild tetraploid bermudagrass Cynodon Dactylon L. using a bacteriophage cloning vector : 2. ligation and in vitro packaging of the recombinant phage DNA molecules

Abstract EN

This paper represent the second step in a molecular cloning program aiming to clone large DNA fragments ()٢ the salt tolerant berf^udagrass (Cynodon dactylon L.) DNA using the bacteriophage (E.MBL3) as a sector.

In this work, a yield of about 100 ng bacteriophage DNA per one liter culture was obtained with.a purity ranging between (٢١٦٠ م(ة.ا-?.ا vcctor DNA was completely double digested with the restriction enzymes BamHI and FcoRl, followed لاظ purification of vector arms.

Bacteriophage arms were also efficiently ligated with the partially digested bermudagrass DNA (prepared earlier).

The optimum ligation ratio of arms: inserts (EMBL3: bermudagrass) was found to be 4:1 respectively.

The recombinant DNA was successfully in vitro packaged and plated on a P2 lysogen E.

coli (strain NM539).

An efficiency of about 1.3xl0r؛ pfu/ ug recombinant phage DNA was determined ١١؛ these experiments.