TY  - ADVS
AU  - Gupta, Tapan
TI  - Molecular Cloning of nodH from Rhizobium meliloti to Develop Escherichia Coli as an Anti - HPV Microbicide
U1  - 660.6 
PY  - 2010///
CY  - Ahmedabad
PB  - Institute of Science, Nirma University
KW  - Biotechnology
KW  - Molecular Cloning
N1  - Abstract: K5 polysaccharide (E. coli K5) having iduronic acid residues are structurally similar to Heparan Sulfate and owing to their structural similarities with HS, K5 derivatives act as an efficient inhibitors of several HSPG-binding molecules, including viral proteins. Heparan sulfate has been shown to be the primary attachment site for HPV; causing cervical cancer. Chemically modified K5 polysaccharide derivatives having different degrees of sulfation has been shown to be an effective inhibitor of HPV pseudovirion particles. Higher degree of sulfation shows more efficient inhibition of HPV. nodH is a gene that catalyses the O-sulfation of the lipochitooligosaccharide(called as =nod factor‘ which helps in the nodulation of the rhizobia to the plant roots) in Rhizobium meliloti 1021. A recombinant plasmid having nodH gene has been constructed for the incorporation into E. coli K5 strain where if it catalyses the O-sulfation of capsule then it will enable the production of heparan sulfate at reduced costs which would be of great medical and industrial significance. E. coli Nissle 1917 is an ideal organism for this purpose as it is a well established probiotic strain and produces K5 capsular polysaccharide. Hence incorporation of nodH gene into this strain could produce heparan sulfate which could be effective in scavenging virus and can be used directly in vivo for scavenging various viral pathogens. This recombinant plasmid was transformed into E. coli DH5a strain. The nodH gene is in proper orientation with respect to the lac promoter. Expression of the clone would be carried out by the detection of protein in E. coli DH5a. O-sulfation activity can be determined in E. coli Nissle 1917 strain by its ability to form heparan sulfate from N-acetylheparosan
ER  -