新西兰白兔CTSB、CTSS基因克隆、生物信息学分析及组织表达研究

doi:10.16431/j.cnki.1671-7236.2023.09.006

Abstract

Abstract: 【Objective】 The CDS regions of cathepsin B (CTSB) and CTSS genes in New Zealand White rabbits were cloned and bioinformatics analysis was performed,and explore the expression regularity in different tissues of New Zealand White rabbits.【Method】 PCR was used to amplify and clone the CDS region of CTSB and CTSS genes,and Mega 7.0 software was used to construct the phylogenetic tree and analyze the amino acid similarity.Bioinformatics software was used to analyze the physical and chemical properties,cross-membrane structures,signal peptides and sub-cellular locations of CTSB and CTSS proteins.The expressions of CTSB and CTSS genes in different tissues of New Zealand White rabbits were detected by Real-time quantitative PCR.【Result】 The sequence length of CDS region of CTSB and CTSS genes in New Zealand White rabbits was 1 020 and 1 023 bp,encoding 339 and 340 amino acids,the molecular weight was 37.627 and 38.253 ku,the theoretical isoelectric point was 5.53 and 8.40,and the instability coefficient was 30.76 and 32.38,which were acidic and basic proteins respectively.The amino acid sequences of CTSB and CTSS genes of New Zealand White rabbits were closely related to pigs and horses,respectively,but were far related to chickens.The secondary structure of CTSB and CTSS proteins was mainly composed of alpha helix and random coil.The predicted tertiary structure of both proteins was consistent with the secondary structure,and both were hydrophilic proteins.CTSB signal peptide splicing sites was located at 17-18 amino acids,containing 1 N-end Propeptide-C1 domain and 1 Peptidase-C1A-C domain,while CTSS signal peptide splicing sites was located at 25―26 amino acids,which contained 1 Inhibitor-129 domain and 1 Peptidase-C1 domain.The results of subcellular localization showed that CTSB and CTSS proteins were mainly distributed in the cytoplasm (including the cell wall).Real-time quantitative PCR results showed that the expression of CTSB gene was higher in abdominal fat and liver,and the expression of CTSS gene was the highest in spleen,both of which were significantly higher than other tissues (P<0.05).【Conclusion】 The full-length CDS sequences of CTSB and CTSS genes in New Zealand White rabbits were successfully cloned,and the expression in different tissues of New Zealand White rabbits were revealed and the biological functions were initially analyzed,which providing a basis and reference for further research on the functions and regulatory mechanisms of CTSB and CTSS genes in rabbits.

Key words: New Zealand White rabbits; CTSB gene; CTSS gene; cloning; bioinformatics analysis; tissues expression

CLC Number:

S829.1

YUAN Dongdong, SONG Guohua, CHEN Mengjuan, WANG Zhitong, BAI Zhongfeng, CAI Hanfang, LI Ming, XU Huifen. Cloning,Bioinformatics Analysis and Tissues Expression of CTSB and CTSS Genes in New Zealand White Rabbits[J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(9): 3499-3509.

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Cloning,Bioinformatics Analysis and Tissues Expression of CTSB and CTSS Genes in New Zealand White Rabbits

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