Cloning and Expression of ClKptA/Tpt1 Gene from Cunninghamia lanceolata（Lamb.）Hook

doi:10.13560/j.cnki.biotech.bull.1985.2020-0177

Abstract

Abstract: Cunninghamia lanceolata（Lamb.）Hook is one of the most important fast-growing timber tree species in southern China. Phosphorus is an essential nutrition in plant growth and development. However,the concentration of phosphorus of soil is relatively low in the subtropical region of China. In order to elucidate how C.lanceolata fully uses phosphorus to promote its growth under phosphorus limitation,the phosphate transferase gene ClKptA/Tpt1 was cloned by RT-PCR from C. lanceolata,and genetic relationship between the protein and KptA/Tpt1 of other species was analyzed. Meanwhile,the tertiary structure of the protein was predicted,and the protein was expressed in Escherichia coli. Furthermore,the expression patterns of ClKptA/Tpt1 in the different parts of C. lanceolata treated with different concentrations of phosphorus were detected. The result showed that the total length of CDS（code sequence）of ClKptA/Tpt1 was 1 143 bp,and it encoded 380 amino acids,which had the closest relationship with the KptA/Tpt1 of Nelumbo nucifera and the furthest relationship with Solanum lycopersicum. The molecular weight of ClKptA/Tpt1 expressed in E. coli was 55.5 kD. The amino acid sequence of ClKptA/Tpt1 did not have a transmembrane structure,and tertiary structure of ClKptA/Tpt1 main constructed by α-heli and β-sheet. ClKptA/Tpt1 expressed in the leaves at the highest and the lowest in stem. The expression level of ClKptA/Tpt1 and lignin content of stem increased with phosphorus content increasingly added in the soil of planted C. lanceolata. These results lay a theoretical foundation for elucidating the molecular mechanism of fully utilizing phosphorus to promote lignin biosynthesis in C. lanceolata.

Key words: ClKptA/Tpt1, affiliation, protein expression, phosphorus treatment, expression pattern

YANG Shi-quan, PENG Dan, FEI Wen-jie, YANG Feng, QU Gao-yi, TANG Wei-wei, OU Jian-ping, DENG Xiang-wen, ZHOU Bo. Cloning and Expression of ClKptA/Tpt1 Gene from Cunninghamia lanceolata（Lamb.）Hook[J]. Biotechnology Bulletin, 2020, 36(8): 15-22.

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