Isolation of development-related genes in somatic embryo radicle of carrot (Daucus carota L.) using modified cDNA RDA |
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Authors: | Fengqiu Diao Lei Zhang Meijuan Huang Naihu Wu |
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Affiliation: | (1) Institute of Developmental Biology, Chinese Academy of Sciences, 100080 Beijing, China;(2) College of Life Sciences, Peking University, 100871 Beijing, China |
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Abstract: | Using modified cDNA RDA capitalizing on the high affinity of streptavidin for biotin and magnetic-absorption-based separation, we have obtained four bands of specifically expressed cDNA in the carrot somatic embryo deregulated for 12 h, which were designated as NR-1, NR-2, NR-3 and NR-4, respectively. As revealed by homology analysis of their DNA sequences after cloning them into pBS, remarkable homology was demonstrated in NR-2, NR-3 and NR-4 with the genes coding for LEA (late embryogenesis abundant protein), Dna J and xyloglucan endo-trans-glycosylase in plants. On the other hand, NR-1 showing no homology with any known sequence may have come from unknown genes. Using 32P-labeled NR-1 as probe, hybridization with cDNA fragment population has shown that we have actually cloned a new gene fragment related to radicle development. As shown by further Southern hybridization, these genes may be present in carrot genome in the form of single or low copies. |
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Keywords: | carrot (Daucus carota L.) cDNA RDA radicle development gene cloning. |
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