This study aims to examine the effect of Genetically Modified β -Carotene Biofortified Rice rice developed by simultaneous expression technology in NAAS on biological immunity. Accordingly, this study added Genetically Modified β-Carotene Biofortified Rice 25, 50% and general rice 50% as control group into diet and provided rats with the prescribed feeds and then measured the contents of immunoglobulin and cytokine in blood. As a result, male and female IgM, IgE, male IgG1, female IgG2a and TNF-a, IL5 and IL12 showed no significant difference; male IgG2a tended to decrease dependently on the combined concentration of Genetically Modified β-Carotene Biofortified Rice; female IgG1 showed significance with control group, but its association with diet was not found. The higher the dietary mixing ratio, the more the male and female IFN-a and female IL-4 contents, regardless of rice variety, and it was found that female IL6 content decreased significantly, but its association with diet was not found. The risk of beta carotene-enriched rice into environment and human body has not been reported yet. The digestion of Genetically Modified β-Carotene Biofortified Rice can be seen as "safe" as this test result showed no big difference between general rice and Genetically Modified β-Carotene Biofortified Rice, and its usability is full of suggestions.
The β-carotene biofortified transgenic rice was developed by transforming rice cv. Nakdongbyeo with phytoene synthase (Psy) and carotene desaturase (Crt I) genes isolated from Capsicum and Pantoea. The aim of this study was to perform molecular characterization of rice transformants of T5-T7 generation harboring Psy and Ctr I genes driven by endosperm specific globulin promoter for biosafety evaluation of β-carotene biofortified transgenic rice. The structure and sequence of T-DNA in the transformation vector and the insertion sites, flanking sequences and generational stability of inserted T-DNA in transgenic rice lines were analyzed. The transformation vector consisted of right border, MAR gene, carotenogenic genes unit, herbicide resistance selectable marker unit, MAR gene and left border in sequential order. T-DNA was introduced at the position of 30,363,938-30,363,973 bp of chromosome No. 2 by adaptor-ligation PCR. Stable integration of T-DNA and stable expression of bar gene was confirmed in T5 to T7 generations. It was also confirmed that the backbone DNA of transformation vector containing antibacterial gene was not present in the genome of β-carotene biofortified transgenic rice. HPLC analysis confirmed that carotenoids were consistently detected through T5-T7 generations.
β-카로틴 강화벼 도입유전자의 도입 위치와 안정성을 도입 위치 주변염기서열 분석과 서던 분석한 결과, 벼 염색체 2번 중 30,363,938번과 30,363,973번 사이의 단일 부위에 도입유 전가 도입되었으며, T5-T7 세대 동안 도입된 모든 유전자들 이 안정적으로 유지되고 있으며, 도입유전자의 운반체인 Backbone DNA (pSB11)는 β-카로틴 강화벼에 삽입되지 않 았음을 확인하였다. 선발 마커로 도입된 PAT 단백질의 발현 분석 결과에서도 T5-T7 세대별, 생육시기별로 안정적으로 발 현됨을 검정하였으며, 목적 형질인 β-카로틴 분석 결과에서 도 모품종인 낙동벼에 비해 9.4배 함량이 증가됨을 확인하였 다. 이상의 분석기법을 통해 복수세대에서 β-카로틴 강화벼 의 도입 유전자들이 안정적으로 유지되고 목적 단백질들이 안정적으로 발현되고 있음을 확인하였다.