Cloning, Tissue Expression and Functional Characteristics of the KITLG Gene of Banna Mini-pig Inbred Line

Purpose In order to study the role of KITLG gene in pig germ cell development.

Methods Based on GenBank KITLG mRNA sequences of pig, the specific primers were designed, and the coding sequences and flanking sequence of KITLG gene were amplified from Banna mini-pig inbred line (BMI) epididymis tissues. Semi-quantitative PCR was applied to analyse the KITLG gene expression profiles of 10 important tissues. KITLG protein sequence was used to carry out functional bioinformatics analysis and construct multiple species phylogenetic tree.

Results The 825 bp coding sequence of BMI KITLG gene was obtained with GenBank accession number KU705624, which encodes a protein of 274 amino acids with GenBank accession number AOC89042. Semi-quantitative PCR in multi-tissues indicated that KITLG gene was expressed highly in the epididymis, heart, spleen, lung, kidney, stomach and low expression in the cerebrum, liver, small intestine and testes. Further functional bioinformatics analysis indicated that KITLG protein contained one conserved domains SCF with a transmembrane region and a signal peptide sequences. Its N-terminal and C-terminal are hydrophilic and its secondary structure is mostly alpha helices. The KITLG protein has five distinct modification sites. Phylogenetic analysis showed that KITLG was highly conserved in evolution and had the closest relationship with cattle and sheep.

Conclusion Our study will lay a foundation for further study of the KITLG gene functions in pig germ cell development.