RESUMEN
AIM: To express and purify the human papillomavirus type 16 E1 protein in E.coli and prepare the antibody against HPV-16 E1. METHODS: HPV-16 E1 gene was amplified by PCR and cloned into prokaryotic expression vector pMAL-p2x, and the recombinant plasmid was transformed into E.coil BL-21. We optimize the soluble expression condition of fusion protein by induction with different IPTG concentration and different temperature. The expressed fusion protein was purified by mahose affinity column Chromatography. To prepare the anti-serum, New Zealand white rabbits were immunized with purified HPV-16 E1 protein via hypodermic and volar. Western blot and ELISA analyzed the serum's specificity against HPV-16 E1 and serum titers. RESULTS: Restriction endonuclease analysis and DNA sequencing showed HPV-16 E1 was cloned into the plasmid pMAL-p2x. Based on the optimization experiments, it concluded that the best soluble expression conditions for the HPV-16 E1 fusion protein involved addition of IPTG to a final concentration of 0.5 mmol/L and then further incubation at 28°C. The purity of the HPV-16 E1 fusion protein was over 95.7% after purification. ELISA and Western blotting showed the titers of the anti-serum were above 1:640 000, and the anti-serum can specifically bind with HPV-16 E1 protein. CONCLUSION: We have ingathered the HPV-16 E1 fusion protein by expressing in E.coli and purifying, and the antibody against HPV-16 E1 was prepared with the fusion protein immunizing New Zealand white rabbits. This work will provide an antigen and detection antibody for further study on the HPV-16 E1 function.
Asunto(s)
Anticuerpos/aislamiento & purificación , Clonación Molecular/métodos , Sueros Inmunes/aislamiento & purificación , Proteínas Oncogénicas Virales/genética , Animales , Anticuerpos/inmunología , Especificidad de Anticuerpos , Escherichia coli , Vectores Genéticos , Humanos , Sueros Inmunes/inmunología , Plásmidos , Conejos , Proteínas Recombinantes/biosíntesis , Proteínas Recombinantes/inmunología , Proteínas Recombinantes/aislamiento & purificaciónRESUMEN
A major diagnostic challenge to the evaluation of an incomplete intestinal obstruction is to distinguish between infectious and malignant etiologies. We present a case of an elderly woman complaining of abdominal pain accompanied with nausea and vomiting, and failure to pass gas or stools. Anti-tuberculosis drugs were used to relieve her abdominal pain, and a needle biopsy of the peritoneal cavity showed evidence of primary papillary serous carcinoma of the peritoneum (PSCP). This is a rare description of tuberculosis in the setting of PSCP. This report illustrates the potential complex nature of malignancies, and emphasizes the need to consider coexistence of malignancy and infection in patients, especially in those with risk factors for malignancy who fail with antibiotic therapy.