Method for rapid screening of bacterial transformants and novel simian adenovirus proteins
AssigneeThe Trustees of the University of Pennsylvania
InventorsGuangping Gao, James M Wilson
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Abstract
Chimpanzee serotype C68 proteins, peptides, and polypeptide are provided. Also provided are novel adenoviruses derived from these proteins, as well as compositions containing these proteins and methods of using same for immunization and therapy. Further, a rapid method for screening recombinant transformants using a visually detectable method is described.
Claims33
A C68 chimpanzee adenoviral capsid protein which is substantially free of other viral proteins with which it is naturally associated, said protein selected from the group consisting of:
- (a) a hexon protein characterized by the amino acid sequence of SEQ ID NO:17;
- (b) a penton protein characterized by the amino acid sequence of SEQ ID NO:
- 12;
- (c) a fiber protein characterized b) the amino acid sequence of SEQ ID NO:27:
- (d) a unique fragment of any of (a) to (c) comprising 8 amino acid residues in length.
A pseudotyped adenovirus comprising a C68 adenoviral capsid having an amino acid sequence as provided in SEQ ID NO:16 encapsidating a molecule comprising adenovirus 5′ inverted terminal repeat sequences (ITR), a minigene, and adenovirus 3′ ITR, wherein each of the 5′ and 3′ ITRs are from a serotype heterologous to C68.
A method for rapid screening of recombinant constructs comprising the steps of:
- (a) providing a first parent construct having a green fluorescent protein (GFP), operably linked to regulatory sequences which permit its expression in a host cell, downstream of the portion of the first parent construct to be in the recombinant construct;
- (b) providing a second parent construct having a GFP operably linked to regulatory sequences which permit its expression in a host cell upstream of the portion of the second parent construct to be in the recombinant construct;
- (c) culturing a selected host cell with the first parent construct and second parent construct under conditions which permits them to homologously recombine;
- and
- (d) screening for the absence expression of GFP expression, which indicates the presence of the recombinant construct.
A host cell comprising:
- (a) a first parent construct having a green fluorescent protein (GFP), operably linked to regulatory sequences which permit its expression in a host cell, downstream of the portion of the first parent construct to be in the recombinant construct;
- and
- (b) a second parent construct having a GFP operably linked to regulatory sequences which permit its expression in a host cell upstream of the portion of the second parent construct to be in the recombinant construct.
Dependencies are inferred from the claim text.