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通过微粒轰击法转化黄瓜组织:鉴定含有功能性和非功能性转移基因的植株。

Transformation of cucumber tissues by microprojectile bombardment: identification of plants containing functional and non-functional transferred genes.

作者信息

Chee P P, Slightom J L

机构信息

Molecular Biology Research Unit, Upjohn Company, Kalamazoo, MI 49007.

出版信息

Gene. 1992 Sep 10;118(2):255-60. doi: 10.1016/0378-1119(92)90196-v.

Abstract

The microprojectile bombardment method was used to transfer DNA into embryogenic callus of cucumber (Cucumis sativus), and stably transformed cucumber plant lines were obtained. A total of 107 independently regenerated cucumber plants were assayed for the presence and expression of the transferred Nos-NPTII gene (encoding nopaline synthase-neomycin phosphotransferase II). Genomic blot hybridization analyses showed that a high percentage (16%) of the cucumber plants were transformed with Nos-NPTII; however, only about 25% of these transgenic plants expressed Nos-NPTII. Inactivity of Nos-NPTII in many of the transformed cucumber plants may be associated with the transfer of multiple copies of Nos-NPTII. PCR and genomic blot hybridization analyses were used to show that the transferred gene was inherited in the subsequent plant generation.

摘要

采用微粒轰击法将DNA导入黄瓜(Cucumis sativus)胚性愈伤组织,获得了稳定转化的黄瓜株系。对总共107株独立再生的黄瓜植株进行检测,以确定转入的Nos-NPTII基因(编码胭脂碱合成酶-新霉素磷酸转移酶II)的存在和表达情况。基因组印迹杂交分析表明,高比例(16%)的黄瓜植株被Nos-NPTII转化;然而,这些转基因植株中只有约25%表达Nos-NPTII。许多转化的黄瓜植株中Nos-NPTII的无活性可能与Nos-NPTII多拷贝的转入有关。采用PCR和基因组印迹杂交分析表明,转入的基因在后续植株世代中得以遗传。

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