RESUMO
Chymosin is widely used in the dairy industry, and much is produced through recombinant DNA in organisms such as bacteria and tobacco. In this study, we used a new transgenic method to express caprine chymosin in corn seeds with lower cost and better storage capability. The recombinant chymosin protein was successfully expressed at an average level of 0.37 mg/g dry weight, which is 0.27% of the total soluble protein in the corn seed. Prochymosin can be activated to produce a chymosin protein with the ability to induce clotting in milk, similar to the commercial protein. The activity of the purified recombinant chymosin was as high as 178.5 U/mg. These results indicate that we have successfully established a technology for generating corn seed-derived caprine chymosin for potential use in the dairy industry.
Assuntos
Quimosina/biossíntese , Vetores Genéticos/química , Plantas Geneticamente Modificadas , Sementes/genética , Zea mays/genética , Agrobacterium tumefaciens/genética , Agrobacterium tumefaciens/metabolismo , Animais , Quimosina/genética , Quimosina/isolamento & purificação , Quimosina/farmacologia , Clonagem Molecular , Ensaios Enzimáticos , Floculação/efeitos dos fármacos , Tecnologia de Alimentos , Expressão Gênica , Vetores Genéticos/metabolismo , Globulinas/genética , Globulinas/metabolismo , Cabras , Cinética , Leite/química , Leite/efeitos dos fármacos , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Regiões Promotoras Genéticas , Proteínas Recombinantes/biossíntese , Proteínas Recombinantes/genética , Proteínas Recombinantes/isolamento & purificação , Proteínas Recombinantes/farmacologia , Sementes/enzimologia , Transformação Genética , Zea mays/enzimologiaRESUMO
OBJECTIVE: To investigate the clinical manifestations and risk factors of the patients from developmental dysplasia of the hip(DDH) family. METHODS: Detailed epidemiology investigation, physical examination, functional movement assessment, lab test and X-ray examination were applied to the whole members of a DDH family. RESULTS: In the family with 9 generations and 218 persons, the incidence of DDH was 31.03% in 145 survivors. Patients mainly manifested bilateral knee and hip joint pain, flexion contracture of hip, limitation in internal and external rotation of hip; a few had arthritic functional disorder, deformation, and limp. The radiography illustrated shallow acetabulum with increased inclination, which encompassed the femoral head badly. Deformation of the femoral head, narrow joint space and osteophyte were also found by X-ray examination. The main risk factors of DDH were genetic factors, gender, birth season etc. The son or daughter with one or two DDH parents had a higher risk for developing DDH than those with no DDH parents. Furthermore, first-degree relatives of the DDH patients also had a greater chance to develop DDH than second-degree relatives and third-degree relatives. The incidence among females was higher than males, and the family member who was given birth in winter had a highest risk for developing DDH. However, there was no difference between incidence of DDH in children and youths and in adults; the incidence of DDH in the immigrants with no blood relationship also did not differ from the incidence of DDH in the family member. CONCLUSION: The genetic factors play an important role in the development of DDH, so do the environmental factors.
