Constructing Eukaryotic Expression Vector and Functional Bioinformatics Analysis of Chicken <i>TNNI2</i> Gene

Jinwei LI; Hongyan SHI; Ming TAN; Shanshan ZHANG; Weiwei ZHANG; Xiaoqing GONG; Jingqiu XIA; Yingning SUN

doi:10.12101/j.issn.1004-390X(n).202008072

JOURNAL OF YUNNAN AGRICULTURAL UNIVERSITY（Natural Science） > 2021 > 36(3): 430-437. > DOI: 10.12101/j.issn.1004-390X(n).202008072

Jinwei LI, Hongyan SHI, Ming TAN, et al. Constructing Eukaryotic Expression Vector and Functional Bioinformatics Analysis of Chicken TNNI2 Gene[J]. JOURNAL OF YUNNAN AGRICULTURAL UNIVERSITY（Natural Science）, 2021, 36(3): 430-437. DOI: 10.12101/j.issn.1004-390X(n).202008072

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

1.
College of Life Science and Agriculture Forestry, Qiqihar University, Qiqihar 161000, China
2.
Miyun District Bureau of Agriculture and Rural Affairs of Beijing Municipality, Beijing 101500, China

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Received Date: August 30, 2020
Revised Date: January 11, 2021
Available Online: May 07, 2021
Published Date: May 30, 2021

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Abstract

Abstract

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.
- eukaryotic expression vector,
- TNNI2,
- preadipocyte,
- bioinformatics analysis

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References (24)

References

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

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