Constructing Eukaryotic Expression Vector and Functional Bioinformatics Analysis of Chicken TNNI2 Gene

PurposeTo construct the eukaryotic expression vector of fast skeletal muscle troponin I (TNNI2) gene, and to transfect and identify in immortalized chicken preadipocyte cell line (ICP1).

MethodsThe RT-PCR was used to clone the whole length of CDS sequence of chicken TNNI2 gene. The eukaryotic expression vector of TNNI2 was constructed by one-step cloning method. The functional bioinformatics analysis was applied to construct the phylogeny tree and study the nucleic acid and the protein of TNNI2.

ResultsThe full-length ORF of TNNI2 was 552 bp, and it encodes 183 amino acids, the theoretical size of the protein was 21.24 ku, the theoretical isoelectric point (pI) was 9.19. Furthermore, the protein had no transmembrane region and no signal peptide region, and was mainly located in the nucleus. It belongs to hydrophilic protein. The protein contains 14 phosphorylation sites, secondary structure of this protein is mainly composed of α-helix. In addition, the TNNI2 gene shares highly homology among different species.

ConclusionWe constructed the pCMV-HA-TNNI2 eukaryotic recombinant vector and transfected it into ICP1 cells successfully, which lays a foundation for further study of TNNI2 function in chicken.