ACS AND ACO GENE CLONING AND PLANT TRANSFORMATION
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    Abstract:

    According to the published sequences, PCR primers of lundnocyclopropane-1-lcarboxylate (ACS) and l-aminocyclopropane-lcarboxylate oxidase (ACO) gene of tomato amplification were designed. Fragments of ACS and ACO gene were generated. The products of PCR fragments and T-vector ligation were transformed into E. colt (DH5a).The recombinants were selected by blue-white colony screening and restriction analysis.After partial sequencing and Southern hybridization, the fragments and stringed products o f two genes were subcloned into plant gene expression vector pBllZI in antisense orientation. The antisense vectors were transformed into Agrobaterium tumefaciens LBA4404 by freeze-thaw method. Fourteendayxild cotyledon pieces were infected with Agrobacterium,and transformants were selected with 75 ig nil-l kanamycin. Southern blotting with 0.gkb fragments of ACS gene as probe provided that two positively transgenic plantletS were obtained.

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王金发,申煌煊. ACS和ACO基因克隆及植物转化[J].热带亚热带植物学报,1998,6(2):105~110

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