Keywords

Spirulina Platensis, Extragenomic Dna, Extraction, Purification, Ctab-Proteinase K

Abstract

Through research, a simple and efficient method for extracting extragenomic DNA (exDNA) from Spirulina – CTAB-proteinase K method was established. In this method, Spirulina cells are first lysed with CTAB extract containing 200 ¦Ìg/ml proteinase K, and then extracted with chloroform/isoamyl alcohol to obtain a crude extract of total DNA. The crude extract is then enzymatically digested with 100 ¦Ìg/ml proteinase K and phenol-containing. /Chloroform/isoamyl alcohol extraction to obtain total DNA, and then use gel freeze-thaw centrifugation to obtain high-purity exDNA. Using the above method, the research results of 6 Spirulina platensis strains Sp-1, Sp-2, Sp-3, Sp-4, Sp-5 and Sp-6 showed that (1) all 6 strains contained exDNA , but the number and relative mass are different; (2) Sp-1, Sp-2, Sp-5 and Sp-6 all have only one kind of exDNA, which is about 1.15kb, 0.75kb, 1.15kb and 1.1kb respectively; ( 3) Both Sp-3 and Sp-4 have 2 exDNA bands. The exDNA of Sp-3 is about 1.55kb and 3.0kb, and the exDNA of Sp-4 is about 1.8kb and 3.6kb; (4) Spirulina species (lines) ) and its relationship with physiological and ecological characteristics, implying that exDNA may be related to major biological issues such as the classification, evolution and morphogenesis of Spirulina, and may be responsible for certain biological functions.

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Original research done by Cao Xuecheng, Wang Zhiping, Yang Lingyong, Xu Bujin

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