| Products/Services Used | Details | Operation |
|---|---|---|
| Gene Synthesis | Vibrio sp. QD-5 was sequenced and the data of the genome were annotated (BerryGenomics, Beijing, China). A cluster of alginate lyase genes was found (Figure 2B), based on which Primers for cloning Aly-IV were designed. The forward primer was designed as 5’- TAGGTGGGATCCGATCATGAAGC-3’ and the reverse primer was designed as 5’-AGAACTCGAGTTGGTACTTACC-3’. The DNA fragments with 1683 bp (Figure 3) were amplified by PCR, and ligated into the vector pET-22b (+) in succession. The resultant plasmids were transformed into E.coil BL21 (DE3) cell. The positive clones were picked up, and the nucleotide sequences inserted into the plasmids were confirmed by DNA sequencing (GenScript, Nanjing, China). | Get A Quote |
The string of bacteria, Vibrio. sp. QD-5 utilizing alginate, was separated from rotten kelp. The results of genome sequencing showed that the strain QD-5 contained four alginate lyase genes.One of the alginate genes Aly-IV was cloned and linked to the plasmid pET-22b (+). The heterologous expressed alginate lyase aly-IVwas characterized，which possessed the theoretical molecular mass of 62 kDa, and theoretical isoelectric point (pI) of 5.12. - The enzyme aly-IV was purified and the activity reached 1256.78 U/mg, with optimal temperature of 35 oC and pH value of 8.9. Nurtured in the temperature below 25 oC for 30 minutes, the activity was almost stable. The result also suggested that the activity of enzyme was ... More
