Agrobacterium 공동배양을 이용한 고추 소포자 유래 초기 배의GFP 발현
GFP expression in the microspore-derived early embryo through co-culturing with Agrobacterium
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The aim of this research is to establish the conditions for Agrobacterium-mediated genetic transformation using microspore. The embryo induction from the microspore was examined under several Kanamycin concentration in media, and the induction rate decreased about 4, 8, 10 times when the Kanamycin concentration increased 10, 50, 100 mg/L, respectively. This indicates that the transformation rate would be much lower if the Kanamycin was used for selection marker. In order to apply the GFP gene as a reporter gene for Agrobacterium-mediated genetic transformation, GFP expression from the microspore- mediated embryos was observed using GFP filter under microscope. The GFP expression occurred when the microspore cultured toward the embryo development for 12, 24 and 48 days. The microspore formed a cluster by microspore division from 12 days culture and continuously became a bigger mass. We obtained a total of 8 GFP-expressing embryos suggesting that the transformation of microspore occurred. However, those young embryos were not fully developed. Further study pertinent to culture conditions is required to fulfill the Agrobacterium-mediated genetic transformation using microspore
Abstract
The aim of this research is to establish the conditions for Agrobacterium-mediated genetic transformation using microspore. The embryo induction from the microspore was examined under several Kanamycin concentration in media, and the induction rate decreased about 4, 8, 10 times when the Kanamycin concentration increased 10, 50, 100 mg/L, respectively. This indicates that the transformation rate would be much lower if the Kanamycin was used for selection marker. In order to apply the GFP gene as a reporter gene for Agrobacterium-mediated genetic transformation, GFP expression from the microspore- mediated embryos was observed using GFP filter under microscope. The GFP expression occurred when the microspore cultured toward the embryo development for 12, 24 and 48 days. The microspore formed a cluster by microspore division from 12 days culture and continuously became a bigger mass. We obtained a total of 8 GFP-expressing embryos suggesting that the transformation of microspore occurred. However, those young embryos were not fully developed. Further study pertinent to culture conditions is required to fulfill the Agrobacterium-mediated genetic transformation using microspore
참고문헌 (9)
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