Construction and Identification of Replicon of Contemporary Zika Virus Strain
Journal Title: Journal of Pharmaceutical and Biomedical Sciences - Year 2017, Vol 7, Issue 5
Abstract
Zika virus (ZIKV) has caused significant outbreaks and epidemics in the Americas recently, and raised global concern due to its ability to cause microcephaly and other neurological complications. In this study, we constructed a subgenomic replicon of ZIKV GZ01, which was a vector containing Renilla luciferase (RLUC) reporter gene, and was Japanese encephalitis pAJE70 as vector. On the basis of this, the RLUC reporter gene sequence was inserted into the coding structure protein gene position instead of the fragment encoding the structural protein (C, prM, E). The ZIKV GZ01 subgenomic replicon containing the RLUC reporter gene was constructed. After the plasmid was linearized, the replicon RNA was transcribed by sp6 promoter and transfected into BHK-21 cells. The replicons were identified by restriction endonuclease digestion with Not I/Kpn I and Hind III and the expression of non-structural protein NS1 was determined by indirect immunofluorescence assay. The luciferase activity was evaluated for viral replication. The results showed that the replicon RNA could express RLUC efficiently in BHK-21 cells, and the RLUC activity increased linearly at 24–48 hours after transfection, which indicated that the replicon had good ability to express foreign genes. The construction of the subgenomic replicon of ZIKV GZ01 strain, which laid the foundation for the study of ZIKV replication and translation mechanism.
Authors and Affiliations
Jiu-Yang Yu, Yu-Qi Yan, Zhen-You Jiang
Keywords
Related Articles
- EP ID EP432773
- DOI -
- Views 88
- Downloads 0
Fibroid Uterus: A Case Study
Uterine fibroids (leiomyomas) are benign smooth muscle tumour that arises within the uterus. By age of 50, 60% of all women will have at least one uterine leiomyoma. The cause of leiomyoma is unknown. They depend on ovar...
Clinical Correlation of Upper Gastrointestinal Endoscopic Biopsies with Histopathological Findings and To Study the Histopathological Profile of Various Neoplastic and Non-Neoplastic Lesions
Background The pathological conditions of upper gastrointestinal tract (GIT) are responsible for a wide range of morbidity and mortality and are also most commonly encountered disorder in routine clinical practice. Endos...
Acute Megakaryoblastic Leukaemia: Experience of Diagnostics at Three Clinical Centres
Background Acute megakaryoblastic leukaemia (AMkL) is a rare hematologic clonal malignancy, characterised by the accumulation of abnormal myeloid blasts with megakaryoblastic differentiation in the bone marrow and/or ext...
Arteriovenous Malformation of the Oral Cavity: A Case Report and Review of Literature
Arteriovenous malformations (AVMs) are developmental vascular anomalies that occur when the embryonic vascular network fails to differentiate. It is composed of abnormal communications between arteries and veins without...
A Proficient Segmentation of Mesenchymal Stem Cells by Curvelet OTSU Particle Swarm Optimisation Technique
Stem cells have authored its own impact in recent times due to their promise in providing newer and unique treatments for a great range of untreatable diseases. This is due to their potential dexterity to rejuvenate and...