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TOOLS OF GENETIC ENGI.

TOOLS OF GENETIC ENGI. By: Russel Jean Gallo. ARG7

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TOOLS OF GENETIC ENGI.

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  1. TOOLS OF GENETIC ENGI. By: Russel Jean Gallo

  2. ARG7 • This marker for nuclear transformation was developed by Saul Purton whilst a post-doc in the lab of Jean-David Rochaix, University of Geneva.  ARG7 encodes argininosuccinatelyase, the last enzyme of the arginine biosynthesis pathway, and the cloned gene can be used to rescue arg7 mutants to arginineprototrophy.  The gene was cloned into the low copy number vector pBR329 to give pARG7.8 [1,2].  We subsequently, inserted a ‘molecular flag’ into one of the introns to allow discrimination between the introducedARG7 DNA and the endogenous arg7 copy.  The flag was a 392 bpHaeII fragment of phiX174 and the resulting plasmid named pARG7.8phi3 [3]. • ARG7 has proved popular as a marker and an insertional mutagen because of the ease of selection and the high transformation rates

  3. BLE • This dominant selectable marker for nuclear transformation confers resistance to the bleomycin family of DNA-damaging antibiotics.  The original version of the marker is described in ref 5, with subsequent improvements in ref 6. 

  4. aphA6 • The marker was developed for chloroplast transformation and confers resistance to kanamycin and amikacin, and is described in ref 7.  Download further details of the plasmid (pSK.KmR) carrying the aphA6 cassette.

  5. Codon Usage Optimizer • CUO is a GUI-based multiplatform software written in Java. It is open source and built with a user/developer-friendly structure. The main function of the software at the current development stage is to optimize genes to be transformed into the Chlamydomonasreinhardtii chloroplast genome although it can be used to optimize genes into other hosts as well. The main tool in CUO, Moptimizer, introduces a semi-automatic way of gene optimization which provides more flexibility and accuracy during the optimization process. The future plan for CUO is to be developed into a multipurpose bioinformatics software where data, findings, planning and learning in biology labs can be created and shared.  CUO can be downloaded using the link below.

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