caa a22 4500 46579534X CHVBK 20180323112036.0 cr unu---uuuuu 170327e19900701xx s 000 0 eng 10.1007/BF00678403 doi (NATIONALLICENCE)springer-10.1007/BF00678403 In-vitro synthesis of functional varicella zoster and herpes simplex viral thymidine kinase [Elektronische Daten] [Ravi Mahalingam, Gary Cabirac, Mary Wellish, Donald Gilden, Abbas Vafai] The varicella zoster virus (VZV) and herpes simplex virus type 1 (HSV-1) thymidine kinase (TK) genes were cloned into the transcription vector pGEM4. In-vitro translation (ivt) of RNA transcribed from these genes showed prominent expression of functional TK proteins with the expected molecular weights of 36 kD for VZV and 43, 39, and 38 kD for HSV-1. The TK proteins were recognized by rabbit anti-VZV and anti-HSV-1 antibodies, respectively. Analysis of the ivt products by thin-layer chromatography revealed the conversion of thymidine to its phosphorylated forms (TMP, TDP, and TTP) by both the VZV and HSV-1 TK genes. The estimated specific activities of the in-vitro translated VZV and HSV-1 TKs were comparable. VZV TK templates were linearized at internal restriction sites and RNAs transcribed from these templates directed the synthesis of polypeptides with sizes consistent with the colinearity of the VZV TK gene. Deletion of 107 amino acids at the carboxy terminus of the VZV TK gene abolished the in-vitro TK activity. In addition, immunoprecipitation of truncated proteins synthesized in vitro suggested the possible involvement of the region between amino acid residues 101 and 168 from the amino terminus of the VZV TK molecule in the formation of structures necessary for antigenicity. Kluwer Academic Publishers, 1990 HSV-1 and VZV thymidine kinase nationallicence in vitro transcription-translation nationallicence Mahalingam Ravi Department of Neurology, University of Colorado Health Sciences Center, 80262, Denver, CO, USA aut Cabirac Gary Department of Neurology, University of Colorado Health Sciences Center, 80262, Denver, CO, USA aut Wellish Mary Department of Neurology, University of Colorado Health Sciences Center, 80262, Denver, CO, USA aut Gilden Donald Department of Neurology, University of Colorado Health Sciences Center, 80262, Denver, CO, USA aut Vafai Abbas Department of Neurology, University of Colorado Health Sciences Center, 80262, Denver, CO, USA aut Virus Genes Kluwer Academic Publishers 4/2(1990-07-01), 105-120 0920-8569 4:2<105 1990 4 11262 https://doi.org/10.1007/BF00678403 text/html Onlinezugriff via DOI 1 research-article jats NATIONALLICENCE 856 40 https://doi.org/10.1007/BF00678403 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Mahalingam Ravi Department of Neurology, University of Colorado Health Sciences Center, 80262, Denver, CO, USA aut NATIONALLICENCE 700 1- Cabirac Gary Department of Neurology, University of Colorado Health Sciences Center, 80262, Denver, CO, USA aut NATIONALLICENCE 700 1- Wellish Mary Department of Neurology, University of Colorado Health Sciences Center, 80262, Denver, CO, USA aut NATIONALLICENCE 700 1- Gilden Donald Department of Neurology, University of Colorado Health Sciences Center, 80262, Denver, CO, USA aut NATIONALLICENCE 700 1- Vafai Abbas Department of Neurology, University of Colorado Health Sciences Center, 80262, Denver, CO, USA aut NATIONALLICENCE 773 0- Virus Genes Kluwer Academic Publishers 4/2(1990-07-01), 105-120 0920-8569 4:2<105 1990 4 11262 Metadata rights reserved Springer special CC-BY-NC licence nationallicence BK010053 XK010053 XK010000 NATIONALLICENCE NATIONALLICENCE NL-springer