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Cloning and Expression Analysis of LmP5CS Gene from Lycium chinense Miller |
FENG Yuan-hang, WANG Gang, JI Jing, GUAN Chun-feng, JIN Chao |
Genetic Engineering Institute of Tianjin University, Tianjin 300072, China |
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Abstract Proline is the important osmotic regulation substances in plants and plays a critical role in improving the stress tolerance of plants. The materials is Lycium chinense Miller,which proline content changes significantly after salt stress. After 1.5% NaCl stress, full length cDNA sequence of a putative Δ1-pyrroline-5-carboxylate synthase gene(P5CS)was cloned from Lycium chinense Miller leaves using RT-PCR and 3’rapid amplification of cDNA ends(RACE),named LmP5CS, and construct expression vector pH7m24GW,3rc -LmP5CS. Sequence analysis showed that the complete open reading frame (ORF) of this gene is 2154 bp, encoding for a protein of 717 amino acids with an isoelectric point of 6.07 and a molecular weight of 77.5 kDa.After 200 mmol/L NaCl stress, protein expression level increased at first and decreased subsequently, proline content changed in accordance with that. The semi-quantitative PCR result suggests that LmP5CS plays an important role in proline content change responses to salt stress.
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Received: 04 September 2012
Published: 25 January 2013
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