Isolation of XXXX PIR genes and their utilization for cell surface display and recombinant protein secretion.

@entity1160 @entity10056 @entity72522 @entity375 @entity43595 @entity2574
Proteins with internal repeats are highly conserved among budding @entity375 . In this study, the isolation of two proteins with internal repeats (PIR) genes, i.e. PpPIR1 and PpPIR2, from the methylotrophic @entity375 @entity10056 has been reported. The @entity72522 and @entity43595 genes' open reading frames were found to contain 1068 and 972 bases, respectively. The sequence homology search showed a homologous conserved repeat of PIR @entity375 block (SQIGDGQIQATT) in both proteins. The PIR @entity375 block was present eight times in the PpPir1p and four times in the PpPir2p proteins. Both proteins showed conserved @entity1160 (Q) and @entity2574 (D) in the repeated sequences, indicating a possible alkali-sensitive b1,3-glucan ester linkage. The fusion constructs of PpPir1-2p and enhanced green fluorescent protein (EGFP) were developed for @entity375 cell surface display. The immunofluorescence assay showed uniform localization of EGFP protein on the @entity10056 cell surface in all fusion constructs. Furthermore, new vectors were developed for recombinant protein secretion in @entity10056 , utilizing the pre-pro signal of PpPir1p protein. Efficient processing of the signal sequence was observed from EGFP and human a1-antitrypsin (AAT) fusion constructs and recombinant protein secretion was obtained in the culture supernatant.
