Address
Malaysia Genome and Vaccine Institute, National Institutes of Biotechnology Malaysia, Jalan Bangi, 43000 Kajang, Selangor, Malaysia
Work Hours
Monday to Friday: 830AM - 530PM
pSRSapI
RM150.00
Purpose / Description: A vector for expressing transgenes in the C. reinhardtiii chloroplast
Reference: New tools for chloroplast genetic engineering allow the synthesis of human growth hormone in the green alga Chlamydomonas reinhardtii
Catalog ID: CC-pSRSapI
Product details
Vector type: Gene expression
Copy number: Unknown
Growth temperature: 37°C
Antibiotics Resistance: Unknown
Gene / Insert: None
Cloning method: SapI and SphI restriction enzymes
Promoter: psaA exon 1 promoter
Resource information:
pSRSapI is a vector for expressing transgenes in the C. reinhardtiii chloroplast (Wannathong et al., 216). The transgene should be inserted into the expression site using SapI and SphI restriction enzymes (or their isoschizomers LguI and PaeI, respectively). It will then be flanked by the C. reinhardtii psaA exon 1 promoter and 5′ UTR (perfect scarless fusion at the ATG start codon) and the C. reinhardtii rbcL 3′ UTR. The external flanking regions in the construct include an intact psbH gene and target the expression cassette downstream of psbH by homologous recombination.
Ampicillin selection in E. coli. Minimal medium for restoration of psbH gene in C. reinhardtii, selecting for phototrophic growth. The recipient cell line must therefore be a psbH mutant such as CC-5168 cw15 psbH [strain TN72].
