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In Vitro Translation of Nodule Specific mRNA from Alfalfa (Medicago sativa L. Vernal) Root Nodules
신평균,김경수,조무제,Shin, Pyung-Gyun,Kim, Kyung-Sao,Cho, Moo-Je 생화학분자생물학회 1986 한국생화학회지 Vol.19 No.2
알팔파근류에 R. meliloti 102F51 및 3A150 mutant ($nif^-$, $nod^+$)를 접종하여 얻은 nodule과 접종하지 않은 뿌리로부터 $poly(A)^+-mRN$를 분리 정제하여 [$^3H$]-leucine 및 [$^{35}S$]-methionine으로 in vitro translation 시킨 후 이들을 leghemoglobin 및 nodule specific protein antibody를 이용하여 immunoprecipitation 시킨다음 2D-polyacrylamide gel 전기영동을 행한 결과 분자량 12,000-30,000 범위내에서 적어도 25종 이상의 근류에서 선택적으로 발현이 촉진되는 단백질들을 확인하였으며 이들은 대두의 경우와 상당한 차이가 있는 것으로 보아 두 과작물간에 이들 단백질 양상이 서로 차이가 있을 것으로 추정된다. Nodule specific proteins (nodulins) were detected by the 2D-polyacrylamide gel electrophoresis of the immunoprecipitated in vitro translation products of the $poly(A)^+-mRN$ A from alfalfa root nodules. At least 25 nodule specific polypeptides in the molecular weight range of 12,000-30,000 were identified in the Rhizobium meliloti 102F51 (wild type) infected alfalfa root nodules. The nodulins accounted for about 40% of [$^3H$]-leucine labeled and 12% of [$^{35}S$]-methionine labeled proteins.
Construction of Rhizobium-E.Coli Shuttle Vector
SHIN, PYUNG GYUN,CHOI, YOUNG JU,YUN, HAN DAE,YANG, MIN SUK,CHO, MOO JE 경상대학교 유전공학연구소 1986 遺傳工學硏究所報 Vol.5 No.-
The vector for gene manipulation of Rhizobium, pGUR9, was constructed by combining the mobilization and broad host range replication function of RSF1010 with E. coli cloning vehicle, pACYC184. The Rhizobium vectors, pGUR15 and pGUR19 were also constructed by combining indigenous plasmid pASR286 and pASR186 from Acacia Rhizobium with pSUP104 and pABR322, respectively. These vectors were stably maintained in a variety of Rhizobium and Agrobacterium. They can be efficiently mobilized by RP4 tra function and have selecton markers and versatile cloning sites of the E. coli vectors. They can be efficiently transfered to Rhizobium by transformation.
알파파 근류에서 선택적으로 발현되는 단백질 mRNA 의 In Vitro Translation
김경수,조무제,신평균 생화학분자생물학회 1986 BMB Reports Vol.19 No.2
Nodule specific proteins (nodulins) were detected by the 2D-polyacrylamide gel electrophoresis of the immunoprecipitated in vitro translation products of the poly(A)^+-mRNA from alfalfa root nodules. At least 25 nodule specific polypeptides in the molecular weight range of 12,000-30,000 were identified in the Rhizobium meliloti 102F51 (wild type) infected alfalfa root nodules. The nodulins accounted for about 40% of [³H]-leucine labeled and 12% of [^(35)S]-methionine labeled proteins.