Tinland B, Schoumacher F, Gloeckler V, Bravo-Angel A M, Hohn B
Friedrich Miescher-Institut, Basel, Switzerland.
EMBO J. 1995 Jul 17;14(14):3585-95. doi: 10.1002/j.1460-2075.1995.tb07364.x.
The VirD2 protein of Agrobacterium tumefaciens was shown to pilot T-DNA during its transfer to the plant cell nucleus. We analyze here its participation in the integration of T-DNA by using a virD2 mutant. This mutation reduces the efficiency of T-DNA transfer, but the efficiency of integration of T-DNA per se is unaffected. Southern and sequence analyses of integration events obtained with the mutated VirD2 protein revealed an aberrant pattern of integration. These results indicate that the wild-type VirD2 protein participates in ligation of the 5'-end of the T-strand to plant DNA and that this ligation step is not rate limiting for T-DNA integration.
根癌土壤杆菌的VirD2蛋白在T-DNA转移至植物细胞核的过程中起到引导作用。我们在此通过使用virD2突变体来分析其在T-DNA整合过程中的参与情况。这种突变降低了T-DNA转移的效率,但T-DNA本身的整合效率不受影响。对用突变的VirD2蛋白获得的整合事件进行Southern分析和序列分析,揭示了一种异常的整合模式。这些结果表明,野生型VirD2蛋白参与了T链5'端与植物DNA的连接,并且该连接步骤对T-DNA整合而言不是限速步骤。