Nur Saidah Said and Helen Susilowati and Deya Karsari and Eryk Hendrianto and Mufasirin and Fedik Abdul Rantam (2014) Kloning Fragmen Gen Non-Struktural 1 (NS1) Virus Dengue Subtipe 1 (DENV-1) Sebagai Kandidat Bahan Vaksin Chimera. Veterinaria Medika, 7 (2). pp. 184-193. ISSN 1979-1305

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Abstract

Dengue infection caused by dengue virus (DENV), which includes family Flaviviridae, which has four serotypes of DENV-1, DENV-2, DENV-3 and DENV-4. Dengue virus has a positive sense RNA genome consisting of three genes encoding structural proteins namely protein C (lipids), the E and M/PRM (glycoprotein) and 7 genes encoding non-structural proteins (NS1, NS2a, NS2b, NS3, NS4a, NS4b and NS5). NS1 protein is a highly conserved glycoprotein that plays an important role in viral replication and maturation, NS1 antigen immunogenicity because it has a high potential, capable of inducing antibodies through binding activity of the complement system. This study aimed to clone and analyze protein NS1 gene encoding the genetic stability of non-structural protein 1 (NS1) of dengue virus subtype 1 (DENV-1) after cloning . The results of this study showed fragments of the gene encoding the NS1 protein of dengue virus subtype 1 (DENV-1) after cloning the DNA plasmid vector (pGEM®-T Easy Vector System) and transformed in E. coli JM109 and then performed the verification testing by PCR followed by electrophoresis showed positive results with the invisibility of amplicon fragments NS1 DENV-1 protein coding genes at 435 bp. After sequencing to look at the stability of the gene fragment encoding the DENV-1 NS1 protein after cloning, there are variations in the composition of which is characterized by the presence of nucleotide insertions, deletions, replacements cloned nucleotide composition compared to the original isolate fragments of the gene encoding the NS1 protein of dengue virus serotype 1 at position initial 1-30 and 425 but the arrangement of positions 31-424 and 426-436 nucleotides stabilized. Keywords : dengue NS1 , cloning , genetic stability

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