[Full-length of human papillomavirus type 18L1 gene optimized using the plant preferred codons and synthesized by overlapping PCR].
OBJECTIVE
The Human papillomavirus type 18L1 (HPV18L1) gene was synthesized by overlapping PCR after optimization using plant preferred codons.


METHODS
The gene sequences of HPV18L1 were obtained from GenBank and analyzed using DNAMAN, Lasergene, Vector NTI and BLAST. The target sequence was selected and modified using plant preferred codons by the Synthetic Gene Designer and JCat (Java Codon Adaptation Tool) with the addition of a His-tag to the C-terminus to construct the full-length modified HPV18L1 (mHPV18L1). mHPV18L1 was divided into 5 large segments, namely LS1 to LS5, with sizes ranging from 204 to 477 bp. Forty-three small oligonucleotide fragments with sizes of 57-59 bp and 6 pairs of primers were designed and synthesized. mHPV18L1 was amplified by overlapping PCR and subcloned into pMD18-T vector. The recombinant plasmid was identified by restriction enzymes digestion and sequencing.


RESULTS
mHPV18L1 was successfully assembled using overlapping PCR. The results of digestion with restriction enzymes and PCR amplification confirmed that the recombinant vector pMD18T- mHPV18L1 contained the inserts with expected size of 1749 bp. mHPV18L1 sequence was confirmed by sequencing.


CONCLUSIONS
mHPV18L1 with plant preferred codons and the recombinant vector pMD18T- mHPV18L1 have been obtained.
Zhao
Ying
Y
Department of Pathogenic Biology, School of Public Health and Tropical Medicine, Southern Medical University, Guangzhou, China. zhaoyingxd@163.com
Zhang
Li-Yi
LY
Liu
Chang-Zheng
CZ
Zhou
Xiao-Hong
XH
D001483:Base Sequence; D036022:Capsid Proteins / Q000235:genetics*; D003001:Cloning, Molecular; D003062:Codon / Q000235:genetics*; D017343:Genes, Plant / Q000235:genetics*; D005813:Genes, Synthetic / Q000235:genetics*; D005822:Genetic Vectors / Q000235:genetics; D052161:Human papillomavirus 18 / Q000235:genetics; D008969:Molecular Sequence Data; D053918:Papillomavirus Vaccines / Q000235:genetics; D016133:Polymerase Chain Reaction / Q000379:methods; D011994:Recombinant Proteins / Q000235:genetics*
D036022:Capsid Proteins; D003062:Codon; D053918:Papillomavirus Vaccines; D011994:Recombinant Proteins; C480565:L1 protein, Human papillomavirus type 18