Cloning of AmPCS Gene from Amaranthus mangostanus and Construction of Its Plant Expression Vectors

doi:10.3969/j.issn.1000-7091.2012.02.009

Abstract

Abstract: The synthesis of phytochelatins (PCs) represents a major heavy metal detoxification mechanism in?higher plants. To contribute to the development of the phytoremediation removing Cd from the soil，a PCS gene was?cloned according to homologous gene conserved region by RT-PCR and RACE method from Amaranthus mangostanus L. ，a typical Cd hyperaccumulator named Tianxingmi. Its full-length cDNA is 1 770 bp and encodes a polypeptide of 485 amino acids with an entire ORF. The AmPCS protein showed high homology to other PCS-like proteins?such as Arabidopsis thaliana、Thlaspi caerulescens、Brassica napus and Brassica juncea between 91. 96% and?95. 67% identity. The AmPCS protein contains a N terminal Pfam:Phytochelatin domain known as a heavy-metal-induced phytochelatin catalysis center and a C terminal Pfam:Phytochelatin_C domain known as a heavy metal?sensor. Therefore，the present results suggested that the PCS gene from Amaranthus mangostanu might be a novel?member of the PCS-like family with the function of phytochelatins synthesis. This gene named AmPCs has been submitted to GenBank with the accession number JN979370. The plant expression vector pBI121 and AmPCS gene with?BamHI 和 SacI site were digested by these two restriction enzymes，were ligated and recombinant，and then plant expression vector pBI-PCS was successfully constructed.?

Key words: Amaranthus mangostanus, Phytochelatins, Cloning, AmPCs, Plant expression vector

CLC Number:

SUN Jing-wen, CHENG Ming-fang, LIANG Guo-qing, WANG Xiu-bin, ZHOU Wei. Cloning of AmPCS Gene from Amaranthus mangostanus and Construction of Its Plant Expression Vectors[J]. ACTA AGRICULTURAE BOREALI-SINICA, 2012, 27(2): 44-49. doi: 10.3969/j.issn.1000-7091.2012.02.009.

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