Journal:

Article Title: Cytomegalovirus Assemblin (pUL80a): Cleavage at Internal Site Not Essential for Virus Growth; Proteinase Absent from Virions

doi: 10.1128/JVI.76.17.8667-8674.2002

Figure Lengend Snippet: Construction of I-site mutant HCMV. (A) HCMV proteinase precursor, its four cleavage sites, and their P3 to P1′ cleavage sequences (1, 55). (B) Amino acid and corresponding nucleotide sequences of the native (left) and mutant (right) I sites, with the new NruI restriction enzyme site underlined. The Ala codon, GCC, was mutated to the Val codon, GTC, by a C-to-T change at nucleotide 115625 (italicized) in the AD169 sequence (6). (C) Homologous recombination between shuttle plasmid EB39 and HCMV-BAC, pHB5, that transfers the A143V-coding mutation into the viral genome to form mutant HCMV-BAC, CKC1. The asterisk denotes a mutation.

Article Snippet: HCMV strain AD169 was from the American Type Culture Collection (no. VR-538; Manassas, Va.).

Techniques: Mutagenesis, Sequencing, Homologous Recombination, Plasmid Preparation

Journal:

Article Title: Cytomegalovirus Assemblin (pUL80a): Cleavage at Internal Site Not Essential for Virus Growth; Proteinase Absent from Virions

doi: 10.1128/JVI.76.17.8667-8674.2002

Figure Lengend Snippet: DNA from I-site mutant virus contains mutagenic nucleotide change. A 1,257-bp sequence beginning at the 5′ end of UL80 was amplified by PCR from five DNA templates and analyzed following incubation without (No Enzyme) or with (+NruI) NruI. Shown is an image prepared from a 1% agarose gel containing the resulting DNA fragments stained with ethidium bromide, together with molecular weight markers (MW, lane 1). PCR templates were wild-type (Wt) and mutant (I−) HCMV-BACs (pBAC; lanes 2 and 7 and lanes 3 and 8, respectively), mutant and wild-type HCMV-BAC virions (BACvir; lanes 4 and 9 and lanes 5 and 10, respectively), and standard AD169 virions (Vir, lanes 6 and 11). Fragment sizes (base pairs) are indicated at the right.

Article Snippet: HCMV strain AD169 was from the American Type Culture Collection (no. VR-538; Manassas, Va.).

Techniques: Mutagenesis, Sequencing, Amplification, Incubation, Agarose Gel Electrophoresis, Staining, Molecular Weight

Journal: Vaccine

Article Title: Novel trimeric human cytomegalovirus glycoprotein B elicits a high-titer neutralizing antibody response

doi: 10.1016/j.vaccine.2018.07.056

Figure Lengend Snippet: Trimeric HCMV gB elicited markedly higher titers of serum HCMV neutralizing activities for fibroblasts. HCMV neutralization titers (IC50), using the MRC-5 fibroblast cell line and HCMV strain AD169wt131, of heat-inactivated (A) and non-heat-inactivated (B) day 63 sera from mice immunized in days 0, 21, and 42 with 1, 5, or 25 μg of trimeric HCMV gB versus Sino gB. HCMV neutralization titers (IC50) of heat-inactivated (C) and non-heat-inactivated (D) day 63 sera from mice immunized with trimeric HCMV gB versus Sino gB using MRC-5 and HCMV strain AD169 (*p < 0.05 compared to the sera from mice immunized with Sino gB in equal amount. **p < 0.05 compared to the sera from mice immunized with 25 μg trimeric HCMV gB).

Article Snippet: HCMV strain AD169 WT131 was provided by Drs. Xiao Wang and Haruhiko Murata (Food and Drug Administration) and strain AD169 was purchased from ATCC.

Techniques: Neutralization