Further evidence for the transformation ofVinca rosea protoplasts byAgrobacterium tumefaciens spheroplasts
Further evidence for the transformation ofVinca rosea protoplasts byAgrobacterium tumefaciens spheroplasts
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根癌农杆菌原生质体转化长春花原生质体的进一步证据
DOI：
10.1007/bf00571299
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发表时间：
1985
影响因子：
6.2
通讯作者：
T. Nagata
中科院分区：
文献类型：
--
作者：
K. Okada;S. Hasezawa;K. Syōno;T. Nagata
按照Hasezawa等人的方法，用根癌农杆菌球体转化玫瑰长春花的原生质体，获得nopalinet型钛质粒。(1981)。这些转化子经常分化管胞，但到目前为止还没有观察到进一步分化为畸胎瘤。以~(14)C-α-酮戊二酸和~3H-精氨酸为底物，提高了酶反应的敏感性和特异性。通过这两种放射性同位素在X线片上的迁移来鉴定诺帕林合成酶的活性。此外，对其中一个转化子的T-DNA结构进行了Southern杂交检测。(1980)，并与长春花进行了比较。
Protoplasts ofVinca roseawere transformed by spheroplasts ofAgrobacterium tumefaciensharboring nopalinetype Ti plasmids according to the procedure of Hasezawa et al. (1981). These transformants frequently differentiated tracheids, but further differentiation to teratomata has not so far been observed. Transformation was confirmed by the improved detection of nopaline synthase, where the sensitivity and specificity of the enzyme reaction was increased by employing14C-α-ketoglutaric acid and3H-arginine as substrates. The nopaline synthase activity was identified by the comigration of these two radioisotopes in the cnromatogram. Furthermore, the T-DNA structure of one of these transformants was examined by Southern hybridization according to Thomashow et al. (1980) and compared with that ofVinca roseacrown gall.