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Codon Optimization> | ). For the present study, the GP nucleotide sequence was codon 153 optimized for human expression (Genscript, Piscataway, NJ) with the exception of the 154 AAGAAGAA sequence, which was left undisturbed. In addition, a derivative of GP was made in 155 which the TMCT region was deleted and replaced by the TMCT region of HPIV1 (Fig. 1C), 156 which was presumptively identified based on hydrophobicity analysis. All inserts were synthetically derived (Genscript) and cloned into the rHPIV1-C∆170 169 antigenomic 170 plasmid containing the unique restriction sites MluI and AscI in the pre-N and N-P positions, 171 respectively, using published methods (17).A double-staining fluorescent plaque assay detecting the co-expression of HPIV1 antigens 207 and EBOV GP was performed to determine the stability of GP expression from all vectors. | Get A Quote |
Mammalian Expression System> | Get A Quote |
The recent 2014-2016 Ebola virus (EBOV) outbreak prompted increased efforts to 20 develop vaccines against EBOV disease. We describe the development and preclinical evaluation 21 of an attenuated recombinant human parainfluenza virus type 1 (rHPIV1) expressing the 22 membrane-anchored form of EBOV glycoprotein GP, as an intranasal (IN) EBOV vaccine. GP 23 was codon optimized and expressed either as a full-length protein or an engineered chimeric form 24 in which its transmembrane and cytoplasmic tail (TMCT) domains were substituted with those of 25 the HPIV1 F protein in an effort to enhance packaging into the vector particle and 26 immunogenicity. GP was inserted either preceding the N gene (pre-N) or between ... More