Abstract
The cytosolic glutathione reductase(GR) gene of chinese cabbage was introduced into Lettuce (Lactuca sativa L.) with particle bombardment method. When cotyledon explants were treated with osmoticum-conditioning medium (0.6 M sorbitol/mannitol) 4 hours prior to and 16 hours after bombardment, it was identified by GUS assay that this condition was the most efficient one for introduction of foreign genes into cotyledon tissue of lettuce with particle bombardment. The stable integration of a GR gene was confirmed by the PCR analysis. 0.3, 0.6, 1.5 kbp PCR fragments of transgenes were obtained by three types of primers designed on the basis of GR sequence. To know whether the expression of the GR gene of pBKs-GR 1 can be stably maintained in the next generation, T$_2$selfing seeds obtained from the transformed mother plants were sowed on MS medium supplemented with 200 mg/L kanamycin sulfate. 70% of seedlings showed resistance to kanamycin.
상추에 particle bombardment를 이용한 배추 Glutathione Reductase (GR)유전자의 형질전환효율 향상 및 형질전환체의 유전분석에 관한 실험을 수행하여 얻은 결과를 요약하면 다음과 같다. Particle bombardment 이용시 사출 전 4시간, 사출 후 16시간 동안 삼투조절제인 0.6M serbitolfmannitol 을 처리하여 형질전환시킨 상추 자엽조직에서 GUS 반응 spot이 가장 많았다. 0.3, 0.6, 1.5 kbp로 나타나는 3가지 종류의 primer를 사용하여 PCR한 결과 각각의 primer에 homology를 타나내는 band를 확인하여 355 promoter와 BcGRl유전자가 삽입이 된 것을 확인하였다. 형질전환된 상추의 자식 종자를 채종하여 kanamycin 200mg/L이 첨가된 MS배지에 파종하여 형질전환시키지 않은 상추의 자식 종자와 비교해 본 결과, 형질전환된 상추의 종자에서 발아된 유묘의 약 70%정도가 kanamycin에 대해 내성을 나타내었다.
