Molecular Cloning and Sequence Analysis on cDNA of Cystatin Gene from Tea Leaves

WANG Zhao-xia; LI Ye-yun; JIANG Chang-jun; YU You-ben

doi:10.13305/j.cnki.jts.2005.03.004

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Journal of Tea Science ›› 2005, Vol. 25 ›› Issue (3) : 177-182. DOI: 10.13305/j.cnki.jts.2005.03.004

Molecular Cloning and Sequence Analysis on cDNA of Cystatin Gene from Tea Leaves

WANG Zhao-xia^{1, 2}, LI Ye-yun¹, JIANG Chang-jun¹, YU You-ben³

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Abstract

Two degenerate primers were designed according to the conserved region among the known plant cystatins. A cDNA fragment of 204βbp was amplified by RT-PCR (reverse transcription polymerase chain reaction) of total RNA extracted from fresh leaves of Tea plant (Camellia sinensis cv Longjing43). A full-length cDNA of the cystatin gene was obtained by 3’/5’RACE (rapid amplification of cDNA ends). The cDNA sequence of this 627βbp clone contained an open reading frame encoding a polypeptide of 101 amino acid residues with a predicable molecular mass of 11.026βKDa. The deduced amino acid sequence contained the motif QXVXG conserved among most members of the cystatin superfamily. By using the program of Blast on GenBank database, the sequence presented a high match with the cystatin genes from other plants, such as European chestnut, Cassava, Cowpea, Tomato, Soybean et al. All researched out sequences were all cystatins, so we can conclude that the cloned sequence is a member of cystatin gene from Tea plant.

Key words

Camellia sinensis / cDNA / clonging / cystatin / sequence

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WANG Zhao-xia, LI Ye-yun, JIANG Chang-jun, YU You-ben. Molecular Cloning and Sequence Analysis on cDNA of Cystatin Gene from Tea Leaves[J]. Journal of Tea Science. 2005, 25(3): 177-182 https://doi.org/10.13305/j.cnki.jts.2005.03.004

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