No abstract available.
Capsid Proteins* ; Capsid*

No abstract available.
Capsid Proteins* ; Capsid* ; Genome*

No abstract available.
Capsid Proteins* ; Capsid* ; Humans* ; Insects*

Capsid ; Capsid Proteins ; Virus Assembly

Capsid Proteins ; genetics ; metabolism ; Simplexvirus ; genetics ; metabolism

OBJECTIVE: To express and purify the antigenic peptide of adeno-associated virus (AAV) capsid conserved regions in prokaryotic cells and prepare its rabbit polyclonal antibody. METHODS: The DNA sequence encoding the conserved regions of AAV capsid protein was synthesized and cloned into the vector pET30a to obtain the plasmid pET30a-AAV-CR for prokaryotic expression and purification of the conserved peptides. Coomassie blue staining and Western blotting were used to identify the AAV conserved peptides. Japanese big ear white rabbits were immunized with AAV conserved region protein to prepare polyclonal antibody, with the rabbits injected with PBS as the control group. The antibody titer was determined with ELISA, and the performance of the antibody for recognizing capsid protein sequences of AAV1-AAV10 was assessed with Western blotting and immunofluorescence assay. RESULTS: The plasmid pET30a-AAV-CR was successfully constructed, and a recombinant protein with a relative molecular mass of 17000 was obtained. The purified protein induced the production of antibodies against the conserved regions of AAV capsid in rabbits, and the titer of the purified antibodies reached 1:320 000. The antibodies were capable of recognizing a wide range of capsid protein sequences of AAV1-AAV10. CONCLUSION We successfully obtained the polyclonal antibodies against AAV capsid conserved region protein from rabbits, which facilitate future studies of AAV vector development and the biological functions of AAV.
Animals ; Antibodies ; Capsid ; Capsid Proteins/genetics* ; Dependovirus/genetics* ; Prokaryotic Cells ; Rabbits ; Recombinant Proteins/genetics*

The RNA genome of poliovirus encodes a long polyprotein precursor and this polyprotein is cleaved proteolytically by viral protease to yield mature proteins. The mature proteins derived from the Pl polyprotein precursor are the component of capsids. To further delineate the process of capsid assembly and encapsidation, in a first attempt, a cell line which expresses the authentic Pl polyprotein was established. CV-1 cells were transfected with the pRCRSVS1P1 plasmid DNA which contains 5'ncr sequences, whole authentic capsid gene of poliovirus and neomycin resistance gene. These cells were treated with G418 for 3 months, and eventually G418 resistant cells were selected and formed colonies. Each colony was picked and grown in the media containing G418. DNA analysis indicated that 1 of 13 neomycin resistant cell lines (R2-18) contains whole poliovirus Pl capsid gene segment which was incorporated into the genome. Immuneprecipitation of cell lysates with sera from rabbit immunized with inactivateded Sabin type 1 particles demonstrated the constitutive expression of the poliovirus Pl capsid protein from R2-18.
Capsid Proteins* ; Capsid* ; Cell Line* ; DNA ; Genome ; Haplorhini* ; Kidney* ; Neomycin ; Plasmids ; Poliovirus* ; RNA

OBJECTIVE: HPV in situ hybridization (ISH) is able to detect HPV DNA and identify integrated HPV DNA by punctuate staining and episomal HPV by diffuse staining in the nuclei. Because the expression of L1 capsid protein disappears after integration of HPV DNA, immunohistochemistry (IHC) of L1 capsid protein can be used as a indirect evidence of integration. Therefore, we tried to evaluate the usefulness of HPV ISH and IHC of L1 capsid protein in Cervical intraepithelial neoplasia (CIN). METHODS: Twenty six cervical lesions from patients with CIN and 19 normal cervical epithelium from patients with leiomyoma were evaluated with HPV ISH and IHC of L1 capsid protein. RESULTS: HPV ISH was positive in 80.8% (21/26) in CIN. Among 21 positive cases, diffuse staining was observed in 42.9% and punctuated and diffuse staining in 57.1%. L1 capsid protein was positive in 65.4% (17/26) of cervical tissue sections and 15.4% (4/26) of cervical smears. The punctuated staining of HPV ISH was correlated with high grade CIN (P=0.007) but expression of L1 capsid protein was not associated with grade of CIN (P>0.05). CONCLUSION: HPV ISH is a useful tool to identify integrated HPV DNA in paraffin-embedded, formalin-fixed cervical tissue. HPV integration confirmed by HPV ISH was associated with high-grade CIN. IHC of L1 capsid protein showed better result using cytology smears than tissue sections.
Capsid ; Capsid Proteins ; Cervical Intraepithelial Neoplasia ; DNA ; Epithelium ; Humans ; Immunohistochemistry ; In Situ Hybridization ; Leiomyoma ; Vaginal Smears

BACKGROUND: Warts are caused by human papilloma viruses (HPVs) infection. Clinically, warts sometimes may be persistent and are recalcitrant to treatments for months or even years. OBJECTIVE: To investigate the relationship between clinical findings, HPV infection and L1 capsid protein. METHODS: Fifty-six samples from patients with cutaneous warts were used in this study. HPV DNA chip analysis and polymerase chain reaction (PCR) were performed for detecting the HPV types. Immunohistochemical stains were done for detecting the expressions of HPV L1 capsid protein. RESULTS: We found different types of HPV infection in warts, as follows; 17 cases of HPV2, 15 cases of HPV27, 8 cases of HPV1, 7 cases of HPV16, 6 cases of HPV4, 5 cases of HPV5, 5 cases of HPV6, 4 cases of HPV11, 3 cases of HPV3 and 3 cases of HPV10. Infection with only one HPV type was identified in 50.0% (28/56) of the investigated samples, whereas concomitant infection with two and three viral types was present in 19.6% (11/56) and 10.7% (6/56) of the cases, respectively. The incidence of concomitant HPV infections found in common warts was significantly more frequent than other clinical types of cutaneous warts. L1 capsid protein was expressed in all of the cutaneous warts. CONCLUSION: Concomitant HPV infections are common in cutaneous warts especially, common warts though the L1 capsid protein were present in all four clinical types. Further studies should be performed to clarify the relationship of recalcitrant to treatment and concomitant HPV infections in cutaneous warts.
Capsid ; Capsid Proteins ; Coloring Agents ; Humans ; Incidence ; Oligonucleotide Array Sequence Analysis ; Papilloma ; Polymerase Chain Reaction ; Warts

Human norovirus (HuNoV) is the major etiological agent of nonbacterial gastroenteritis worldwide. However, due to the absence of a rapid and sensitive diagnostic system, detection and monitoring have been limited. The HuNoV genome is composed of three open reading frames (ORFs). And major capsid protein, ORF2, is designated as a viral protein 1 (VP1). In this study, the baculovirus expression system was used for expression of the HuNoV capsid protein, VP1. Recombinant baculoviruses can be used as potent tools in HuNoV studies.
Baculoviridae ; Capsid ; Capsid Proteins ; Gastroenteritis ; Genome ; Humans ; Norovirus ; Open Reading Frames