Characterization of Natural Plasmid and Construction of Putative Transformation Vector Using the Plasmid in Korean Red Alga, Porphyra tenera

Algae > Volume 16(3); 2001 > Article

Algae 2001;16(3): 287-294.

Hack Sun Choi, Kyong Hee Choi, Tae Hoon Kim, Choon Hwan Lee, Tae Hyong Rhew

Dept. of Biology, Dept. of Molecular Biology, Pusan National University

ABSTRACT

Four circular DNA plasmids which were designated as PT1, PT2, PT3, and PT4, respectively, were isolated from marine red macroalga, Porphyra tenera. Among them, the PT4 was cloned by introducing Eco RV digested-linear PT4 into the pBluescript ⅡKS+ vector. The complete DNA sequence of the circular PT4 plasmid was characterized. It contained 1808 bp and had two inverted repeat (20 nucleotides) along with short tandem repeat (12 nucleotides). The plasmid had one open reading frame that could encode a 134 amino acid polypeptide. In an attempt to construct a transformation and expression vector for a marine macroalgae, a plasmid pPTH-smGFP was constructed with the cloned recombinant plasmid, pPT4. The non-destructiveness of the assay method for the green fluorescence protein could allow the privilege of the constructed vector as an efficient and economical expression vector, which may provide an affirmaive way of visual selection of transgenic cells.

Key words: natural DNA plasmid, Porphyra tenera, red alga