Improvement of the aluminum borate whisker-mediated method of DNA delivery into rice callus
Improvement of the aluminum borate whisker-mediated method of DNA delivery into rice callus
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DOI：
10.1626/pps.7.45
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发表时间：
2004-03-01
影响因子：
2.5
通讯作者：
Tanaka, O
中科院分区：
文献类型：
--
作者：
Mizuno, K;Takahashi, W;Tanaka, O
我们改进了硼酸铝晶须介导的DNA导入水稻愈伤组织的方法，CV. Notohikari）。考察了以下因素：晶须的量、搅拌装置的种类、晶须的类型、搅拌的持续时间和搅拌速度。在含有硼酸铝晶须（ABW）、液体培养基和携带β-葡萄糖醛酸酶（GUS）基因的质粒pAct 1-F的1.5ml微管中搅拌20个愈伤组织的盾片组织。搅拌后，对盾片组织进行瞬时GUS表达测定，其将GUS表达细胞可视化为蓝色斑点。将DNA递送的效果评价为由测定产生的蓝色斑点的数量。在本研究中，我们成功地提高了DNA输送的效率，改变搅拌装置从旋转（涡流精灵2）多方向（MT-360），和ABW的类型从Alborex Y氨基硅烷涂层Alborex（Alborex YS 3A）。
We improved the aluminum borate whisker-mediated method of DNA delivery into rice callus (Oryza saliva L., cv. Notohikari). The following factors were examined: amount of whiskers, the kind of apparatus for agitation, the type of whiskers, duration of agitation, and agitation speed. Twenty callused scutellar tissues were agitated in a 1.5 ml microtube containing aluminum borate whiskers (ABW), liquid medium, and the plasmid pAct1-F carrying the beta-glucuronidase (GUS) gene. After the agitation, the scutellar tissues were subjected to transient GUS expression assay, which visualizes GUS-expressing cells as blue spots. Efficiencies of DNA delivery were evaluated as the number of blue spots resulting from the assay. In the present study, we succeeded in improving the efficiency of DNA delivery by changing the apparatus for agitation from rotary (Vortex Genie 2) to multidirectional (MT-360), and the type of ABW from Alborex Y to an aminosilane-coated Alborex (Alborex YS3A).