pHGN3

Restriction digests of the clone give the following sizes (kb): BamHI–4.2; EcoRI–4.2; SalI/HindIII–3.2, 1.0. (ATCC staff) Shuttle vector component of a virulence tester system to be used with the avirulent rabbit poxvirus deletion mutant RPmuhr23HA+ (ATCC VR-2379). [2] DNA cloned into the MCS is integrated into VR-2379 by homologous recombination, possibly altering the host range and virulence of the virus.[2] The hemagglutinin insert is a 1.8 kb 5’SalI/3’HindIII fragment containing approximately 443 bp 5′ flanking sequence, 988 bp coding sequence interrupted by a polylinker, and 408 bp 3′ flanking sequence. [2] The gene is interrupted at the AccI site (nt 43 of the coding region) by a 47 bp MCS from pUC9. [2] Constructed by deleting lacZ and the polylinker from pUC9 by a HaeII digest. [2] The order of the major features in the plasmid is: pUC9 delta lacZ – KpnI – 5′ end of insert – PstI/MCS/EcoRI – 3′ end of insert – KpnI.[2] Recombinants can be identified by inactivation of the hemagglutinin gene (HA-). [2] Medium is 1227 LB plus ampicillin. RPV = rabbit pox virus, vaccinia virus NCBI gi: 335806 Hosts: E.coli, vertebrate cells, chicken embryo fibroblasts virus-infected.