| Production of recombinant proteins GST L1, E6 and | | | | technology in order to undergo serological study in |
| E7 tag HPV 16 to be used in LUMINEX assay for | | | | Tunisian female population. So, HPV types 16 L1, E6 |
| antibody detection among Tunisian female population | | | | and E7 sequences fused to their 3’-end to a |
| with cervical cancer and controls | | | | sequence encoding the terminal undecapeptide of the |
| Achour. M*a, Ben Younes. Ra, Kahla. Sa, Kochbati. Lb, | | | | SV40 large T-antigen (tag) were isolated from |
| Zeghal. Dc, Maalej. Mb, Zouari. Fc, Oueslati. Raa- | | | | plasmids and inserted into a pGEX vector for |
| Laboratory of Environmental Immuno-Microbiology | | | | expression as GST fusion proteins in E.coli. Coding |
| and cancerogenesis (IMEC Unit) Faculty of Sciences | | | | sequences for L1tag, E6tag and E7tag of HPV 16 |
| Bizerta 7021 Zarzouna Tunisia.b- Service of | | | | respectively were mobilized by digestion with |
| Radiotherapy Institute of Cancer Salah Azeiz | | | | enzymes and ligated into digested plasmids |
| Tunisia.c- Service of Gynecology- obstetrics Center | | | | downstream of the GST domain. An expression |
| of Maternity and Neonatalogy Hospital La Rabta | | | | plasmid for GST tag was constructed by inserting a |
| Tunisia. | | | | fragment coding for the tag epitope. Cells of E.coli BL |
| *Corresponding author | | | | 21 were transformed with the pGEX plasmids and |
| Achour Mongia | | | | grown in Luria Bertani medium containing the ampicillin. |
| E-mail: mahaachour2000@yahoo.fr | | | | Recombinant proteins expression was induced by |
| Tel: 216 72 591 845 | | | | adding 0.25 mM isopropyl-?-D-thio-galactoside (IPTG) |
| Fax: 216 72 590 566 | | | | to the medium. The bacteria were harvested after |
| ABSTRACT: | | | | induction and pelleted bacteria were resuspended in |
| Certain types of Human Papillomavirus (HPVs), mainly | | | | phosphate buffered saline (PBS) and lysed using a |
| HPV types 16 and 18 have been recognised as major | | | | high- pressure homogenizer. Lysates were then |
| etiological factors for the development of cervical | | | | cleared by centrifugation. |
| cancer. Antibodies against HPV antigens have been | | | | Proteins were verified by migration in sodium dodecyl |
| found to be associated with cervical cancer evolution. | | | | sulphate (SDS) gel electrophoresis. Data showed that |
| Different assays can be used for antibody detection | | | | they were stable for detection and these lysates |
| but they have low levels of sensitivity and specificity. | | | | have been conserved at -20°C to be used in |
| In the present work we are interested to prepare | | | | Luminex for detection of antibodies in female Tunisian |
| recombinant proteins to be used in LUMINEX | | | | population. |