A Simple and Rapid PCR-based Procedure for Identifying Homozygous Transgenic Rice Plants |
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Authors: | Zheng-Ge Zhu Ray Wu |
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Institution: | (1) College of Life Sciences, Hebei Normal University, Shijiazhuang, Hebei, 050016, People’s Republic of China;(2) Department of Molecular Biology and Genetics, Cornell University, Ithaca, NY 14853, USA |
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Abstract: | We present a simple and rapid method for screening second-generation transgenic rice plants (T1) to identify homozygous plants. The plasmid (pfd11) used for rice transformation contains a partially deleted cytochrome
c gene (cyc) for comparing with the endogenous cyc for copy number. After polymerase chain reaction (PCR) amplification of a segment
of the cyc in transgenic rice DNA followed by agarose gel electrophoresis, two specific bands are obtained. The upper band
represents the endogenous cyc, and the lower band represents the partially deleted cyc in the transgene. The first-generation
plants (T0) that harbor a single copy of the transgene are selected based on the fact that the density of the lower band is half as
dense as the upper band. Next, only plants harboring a single copy of the transgene are advanced to the second generation
(T1). The same PCR procedure is used again, and homozygous T1 plants are easily identified from samples in which the intensity of the two bands is the same. |
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Keywords: | Homozygous plants Rapid PCR analysis Transgenic rice |
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