형질전환 베타카로틴 강화 콩 계통 선발 및 도입유전자 특성 분석
Selection of β-carotene Enhanced Transgenic Soybean Containing Single-copy Transgene and Analysis of Integration Sites
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초록
국내 품종인 광안콩을 이용해 베타카로틴 성분이 강화된유전자변형 콩이 최근 개발되었다. 본 연구에서는 다수의 도입유전자가 염색체에 삽입된 유전자변형 콩을 후대 육성하여유전자분리를 통해 single T-DNA가 도입된 후대계통을 선발하여 작물재료를 육성하고, 또한 도입유전자 삽입위치를 분석하여 GMO 안전성평가를 위한 기초자료로 활용하고자 하였다. 형질전환 콩의 후대 계통에 대한 Southern blot 분석으로4계통의 단일 도입유전자가 도입된 형질전환 계통을 선발하였다. 도입위치 분석결과 3계통(7-1-1-1, 9-1-2, 10-10-1)에도입된 T-DNA는 염색체의 intergenic 부위에 삽입되어 있는반면, 10-19-1 계통의 T-DNA는 14번 염색체내의 유전자exon 부위에 삽입된 것으로 분석되었다.
Abstract
The β-carotene biofortified transgenic soybean was developed recently through Agrobacterium-mediated transformation using the recombinant PAC (Phytoene synthase-2A-Carotene desaturase) gene in Korean soybean (Glycine max L. cv. Kwangan). GM crops prior to use as food or release into the environment required risk assessments to environment and human health in Korea. Generally, transgenic plants containing a copy of T-DNA were used for stable expression of desirable trait gene in risk assessments. Also, information about integration site of T-DNA can be used to test the hypothesis that the inserted DNA does not trigger production of unintended transgenic proteins, or disrupt plant genes, which may cause the transgenic crop to be harmful. As these reasons, we selected four transgenic soybean lines expressing carotenoid biosynthesis genes with a copy of T-DNA by using Southern blot analysis, and analyzed the integration sites of their T-DNA by using flanking sequence analysis. The results showed that, T-DNA of three transgenic soybean lines (7-1-1-1, 9-1-2, 10-10-1) was inserted within intergenic region of the soybean chromosome, while T-DNA of a transgenic soybean line (10-19-1) located exon region of chromosome 13. This data of integration site and flanking sequences is useful for the biosafety assessment and for the identification of the β-carotene biofortified transgenic soybean.
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