DNA shuffling of the GP3 genes of porcine reproductive and respiratory syndrome virus (PRRSV) produces a chimeric virus with an improved cross-neutralizing ability against a heterologous PRRSV strain.
Porcine reproductive and respiratory syndrome virus (PRRSV) is an important swine pathogen. Here we applied the DNA shuffling approaches to molecularly breed the PRRSV GP3 gene, a neutralizing antibodies inducer, in an attempt to improve its heterologous cross-neutralizing ability. The GP3 genes of six different PRRSV strains were bred by traditional DNA shuffling. Additionally, synthetic DNA shuffling of the GP3 gene was also performed using degenerate oligonucleotides. The shuffled-GP3-libraries were cloned into the backbone of a DNA-launched PRRSV infectious clone pIR-VR2385-CA. Four traditional-shuffled chimeras each representing all 6 parental strains and four other synthetic-shuffled chimeras were successfully rescued. These chimeras displayed similar levels of replication both in vitro and in vivo, compared to the backbone parental virus, indicating that the GP3 shuffling did not impair the replication capability of the chimeras. One chimera GP3TS22 induced significantly higher levels of cross-neutralizing antibodies in pigs against a heterologous PRRSV strain FL-12.
Department of Biomedical Sciences and Pathobiology, College of Veterinary Medicine, Virginia Polytechnic Institute and State University (Virginia Tech), Blacksburg, VA 24061-0913, USA.
Lei
L
Zhou
Yan-Yan
YY
Ni
Pablo
P
Piñeyro
Brenton J
BJ
Sanford
Caitlin M
CM
Cossaboom
Barbara A
BA
Dryman
Yao-Wei
YW
Huang
Dian-Jun
DJ
Cao
Xiang-Jin
XJ
Meng
D000595:Amino Acid Sequence; D000818:Animals; D057134:Antibodies, Neutralizing / Q000276:immunology*; D000914:Antibodies, Viral / Q000276:immunology*; D001483:Base Sequence; D002460:Cell Line; D003429:Cross Reactions; D032621:DNA Shuffling; D004195:Disease Models, Animal; D005838:Genotype; D008969:Molecular Sequence Data; D009500:Neutralization Tests; D010802:Phylogeny; D019318:Porcine Reproductive and Respiratory Syndrome / Q000821:virology; D019316:Porcine respiratory and reproductive syndrome virus / Q000145:classification / Q000235:genetics* / Q000276:immunology* / Q000502:physiology; D013552:Swine; D019562:Viral Load; D015678:Viral Structural Proteins / Q000187:drug effects* / Q000276:immunology*; D014779:Virus Replication
D057134:Antibodies, Neutralizing; D000914:Antibodies, Viral; D015678:Viral Structural Proteins