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Isolation and Characterization of Copper- and Zinc- Binding Metallothioneins from the Marine Alga Ulva compressa (Chlorophyta).

Int J Mol Sci. 2019; 
Zúñiga A1,2, Laporte D1, González A1, Gómez M1, Sáez CA2,3, Moenne A1.
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Codon Optimization UcMTs were synthesized and subjected to codon optimization for expression in E. coli by Genscript (Piscataway, NJ, USA) and then ligated to the expression vector pGEX-5X-1 (Genscript) which allowed fusion of UcMTs ORFS with the enzyme glutahione-S-transferase (GST) from the platyhelminthes Schistosoma japonicum (26 Kda), an enzyme containing a single cysteine that does not bind metals; the fusion proteins were named UcMT1.1-GST, UcMT2-GST, and UcMT3-GST. The recombinant vectors were sequenced by Genscript to verify the correct insertion of complete ORFs. Get A Quote

摘要

In this work, transcripts encoding three metallothioneins from Ulva compressa (UcMTs) were amplified: The 5'and 3' UTRs by RACE-PCR, and the open reading frames (ORFs) by PCR. Transcripts encoding UcMT1.1 (Crassostrea-like), UcMT2 (Mytilus-like), and UcMT3 (Dreissena-like) showed a 5'UTR of 61, 71, and 65 nucleotides and a 3'UTR of 418, 235, and 193 nucleotides, respectively. UcMT1.1 ORF encodes a protein of 81 amino acids (MW 8.2 KDa) with 25 cysteines (29.4%), arranged as three motifs CC and nine motifs CXC; UcMT2 ORF encode a protein of 90 amino acids (9.05 kDa) with 27 cysteines (30%), arranged as three motifs CC, nine motifs CXC, and one motif CXXC; UcMT3 encode a protein of 139 amino acids (13.4 kDa) with... More

关键词

Ulva compressa; copper and zinc; expression in bacteria; marine alga; metal accumulation; metallothioneins
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