水稻T-DNA突變種原庫之建立與營運

Abstract

基因功能消除或基因功能活化法是硏究基因功能最直接的路徑，以T-DNA插入基因產生突變，為最普遍被用來消除或活化基因功能的方法。為了評估需要多少個品系才能使每個基因皆可T-DNA插入，必須考慮三個決定T-DNA插入機率的因子：基因的大小、基因組的大小及T-DNA插人的數目，以水稻而言預估大約需要246,000個T-DNA插入品系才能使基因組達到飽和突變。目前中央研究院利用T-DNA插入基因突變法生產出了大量的轉殖水稻株，並進行T-DNA插入點二翼的核酸序列解讀，建立被插入核酸序列資料庫，結合農藝性狀的資料，成立水稻功能性基因的網站，以供研究人員搜尋有興趣的基囚以及其T-DNA插入的品系。這些水稻T-DNA突變株的種子，將放置於國家作物種原中心內保存，以確保種子的品質和壽命。種子的入庫依中央研究院轉基因植株的生產速度而定，預估每年可人庫20,000份材料，依此5年內約可完成100,000份的材料入庫貯藏。此外本中心亦進行突變水稻種子的電腦編號，以便使水稻基因插入部位核酸序列之資料與突變種子庫有一致之電腦編號，方便使用者上網查詢資料與索取種子。這些種子將成為研究水稻功能性基因的重要材料。 The technologies of gene knockout and activation tagging provide a direct route to determining the function of a gene. T-DNA insertion mutagenesis is one of the most popular methods for understanding gene function. Three variables, namely the size of the gene, the size of the genome and the number of T-DNA inserts, were used to determine the probability that a T-DNA insert will be found within a given gene distributed amongst the population. It is estimated that about 246,000 T-DNA transfomed lines are needed to saturate the mutations of rice genome. Transgenic rice carrying T-DNA are being generated at Academia Sinica. The rice genetic DNA flanking T-DNA will be analyzed and the phenotypes of the gene knockout and/or activation tagging mutants will be analyzed. An inserted T-DNA rice website (hitp://trim.sinica.edu.tw) contains the flanking DNA database and the phenotypes that have been evaluated for researchers in Taiwan. The transgenic rice seeds carrying T-DNA will be conserved in the National Plant Genetic Resources Center (NPGRC) of the Taiwan Agricultural Research Institute to ensure their quality and viability. We intend to store 100,000 accession of rice T-DNA tagged line at the NPGRC in the next five years, according to the production rate of 20,000 lines that Academia Sinica is generating each year. NPGRC will entrust file numbers of the transgenic seeds to computer and the seed stock will also be posted on a website. These seeds will be distributed to scientists for phenotypic screening in the near future, which will provide important and precious materials for rice functional genome research in Taiwan.