Construction of Novel Phytochelatins by Overlap Oligonucleotides

Authors

  • Behnaz Saffar Genetic Department, Faculty of Science and Research Institute of Biotechnology, Shahrkord University, Shahrkord, Iran.
  • Mohsen Mobini Genetic Department, Faculty of Science and Research Institute of Biotechnology, Shahrkord University, Shahrkord, Iran.
  • Azadeh Mohamadi Genetic Department, Faculty of Science and Research Institute of Biotechnology, Shahrkord University, Shahrkord, Iran.

Keywords:

Design of oligonucleotide, Hexahistidine, Synthetic phytochelatin

Abstract

Synthetic phytochelatins are protein analogs of phytochelatin with similar heavy metal binding affinities that can be easily produced from a synthetic DNA template. We design synthetic phytochelatin [(Glu-Cys)n Gly] linked to hexahistidine by viral linker peptide and then followed by gene synthesis and cloning of it. Then peptide coding gene (synthetic phytochelatin with linker and hexahistidine) was designed exactly and constructed with step by step methods by overlapping oligonucleotides using T4 DNA Ligase. Finally, synthesized gene amplified by PCR, cloned in pTZ57R/T and transformed to Escherichia coli (DH5α). The results of sequencing show that some types of synthetic phytochelatin (EC4, EC12, and EC20) with linker and hexahistidine were constructed and cloned in vector.

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References

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Published

01-07-2012

How to Cite

Saffar, B., Mobini, M., & Mohamadi, A. (2012). Construction of Novel Phytochelatins by Overlap Oligonucleotides. Journal of Advanced Laboratory Research in Biology, 3(3), 135–138. Retrieved from https://e-journal.sospublication.co.in/index.php/jalrb/article/view/114

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