Cloning,expression of<Emphasis Type="Italic">Crocus sativus</Emphasis> phytoene desaturase gene and preparation of antiserum against it |
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Authors: | Bai?Jie Miao?Chen Xu?Ying Tang?Lin Wang?Zhi-tao Email author" target="_blank">Chen?FangEmail author |
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Institution: | (1) College of Life Sciences, Sichuan University, 610064 Chengdu, Sichuan, China |
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Abstract: | A 2 149 bp full length phytoene desaturase (PDS) cDNA was first cloned from saffron (Crocus sativus L.) stigma using RT-PCR technique and a rapid amplification of cDNA end (RACE) strategy. The cDNA has an open reading frame
of 1 697 bp, which encodes a polypeptide of 565 amino acids. The coding region of the cDNA was inserted into a prokaryotic
expression vector pET-21a(+) and over-expressed inE. coli BL21 (DE3). The fusion proteins were found largely in an insoluble inclusion bodies. The purified fusion protein was used
to immunize rabbits to obtain polyclonal antiserum with titer of 1×105. Western blot analysis by using this particular antiserum showed that the higher expression level of PDS in mature stigma
than in leaves and stamen, and the higher expression level of PDS in mature stigma than in young stigma.
Foundation item: Supported by the Doctoral Foundation of the Ministry of Education, P. R. China and the Young Science Foundation of Sichuan
University (Grant 0020405505012)
Biography: Bai Jie (1968-), female, Ph. D candidate, research direction: plant developmental biology and reproductive engineering. |
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Keywords: | saffron carotenoids phytoene desaturase gene expression antiserum Western blot |
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