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Characterization, and expression profile of a phenylalanine ammonia lyase gene from Jatropha curcas L.

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Abstract

A PAL gene designated as JcPAL1 was cloned from J. curcas L. The full-length is 2336 bp in size with one intron and two exons, encoding a polypeptide of 713 amino acids. Its 5′-upstream region is rich in putative cis-elements including not only PAL typical TATA box, L-box and transcriptional initiation site (TIS) but also light responding motifs. Expression pattern analysis indicated that JcPAL1 were expressed in all tissues, most highly in flowers. When Treated with ABA, GA3, high and low temperature, expression of JcPAL1 were induced. Recombinant JcPAL1 has a pH optimum at 8.7 and a temperature optimum at 60°C in 100 mM Tris–HCl buffer. The Km and Kcat values are 0.125 mM and 1.73 S−1 for l-phenylalanine, and 1.312 mM and 0.109 S−1 for l-tyrosine, respectively. These findings suggested that JcPAL1 might involve in the J. curcas responding to various stresses and l-Phe should be its true physiological substrate. This study is essential prior to uncover whether and how the PAL initiated phenylpropanoid metabolic networks functioning in the defense responses of J. curcas.

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Abbreviations

CHS:

Chalcone synthase

CTAB:

Cetyltrimethylammonium bromide

Inr:

Initiator

IPTG:

Isopropyl-β-d-thiogalactopyranoside

MIO:

4-Methylidien-imidazole-5-one

ORF:

Open reading frame

PAL:

Phenylalanine ammonia lyase

RACE:

Rapid amplification of cDNA ends

RT-PCR:

Reverse transcription-polymerase chain reaction

4CL:

4-Coumarate CoA ligase

SDS-PAGE:

Sodium dodecyl sulfate polyacrylamide gel electrophoresis

TIS:

Transcriptional initiation site

Tris:

Tris(hydroxymethyl) aminomethane

UTR:

Untranslated region

E. coli :

Escherichia Coli

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Acknowledgments

This research is supported by National Key Technology R&D Program of 11th Five-Year Plan of China (No. 2006BAD07A04), General Program of National Natural Science Foundation of China (No. 30670204), China International Science and Technology Cooperation Project (No. 2006DFB63400).

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Authors and Affiliations

  1. School of Life Sciences, Sichuan University, 24 South Section 1, 1st Ring Road, Chengdu, 610065, People’s Republic of China

    Jihai Gao, Shuwen Zhang, Feng Cai, Xiaojiang Zheng, Na Lin, Xiaobo Qin, Yangchao Ou, Xiaoping Gu, Xihong Zhu, Ying Xu & Fang Chen

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  1. Jihai Gao
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Correspondence to Ying Xu.

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Jihai Gao and Shuwen Zhang are Co-first authors.

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Gao, J., Zhang, S., Cai, F. et al. Characterization, and expression profile of a phenylalanine ammonia lyase gene from Jatropha curcas L.. Mol Biol Rep 39, 3443–3452 (2012). https://doi.org/10.1007/s11033-011-1116-4

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