Abstract: This research aims at studying the F pathogen gene of Newcastle disease from the angle of molecular biology, exploring the genetic variation and protein structure of（Newcastle Disease Virus, NDV）F gene. The RNA genome of NDV La Sota strain was extracted, Primer5.0 analysis software was utilized to design primers according to the sequence of NDV La Sota strain published on the GenBank. F gene was amplified and cloned to the pMD18-T vector. The recombinant plasmid pMD-NDV-F was identified by restriction enzyme analysis, PCR identification and nucleotide sequence analysis. Boinformatics software was used to analyze F gene on phylogenetic tree analysis, amino acid sequence homology analysis, transmembrane region analysis and the tertiary structure prediction of F protein. The sequencing results revealed that F gene was 1792 bp and encoded 553 amino acids. The analysis of bioinformatics indicated that F gene was conserved among species and was homologous with NDV La Sota, Clone30, V4, B1 and other weak strains with typical attenuated strain characteristics. The highest amino acid homology of F gene to NDV La Sota strain F gene can reach 100%. The tertiary structure prediction results manifested that the similarity between partial fragment of F protein and NDV F protein fusion fragment was 96%. The study may lay a foundation for the molecular virology research on NDV.