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Molecular cloning and characterization of different expression of MYOZ2 and MYOZ3 in Tianfu goat.

ABSTRACT: The myozenin family of proteins binds calcineurin, which is involved in myocyte differentiation of skeletal muscle. Moreover, gene expression of myozenin is closely related to meat quality. To further understand the functions and effects of myozenin2 (MYOZ2) and myozenin3 (MYOZ3) genes in goat, we cloned them from Tianfu goat longissimus dorsi muscle. Sequence analyses revealed that full-length coding sequence of MYOZ2 consisted of 795 bp and encoded 264 amino acids, and full-length coding sequence of MYOZ3 consisted of 735 bp and encoded 244 amino acids. RT-qPCR analyses revealed that mRNA expressions of MYOZ2 and MYOZ3 were detected in heart, liver, spleen, lung, kidney, leg muscle, abdominal muscle, and longissimus dorsi muscle. Particularly high expression levels of MYOZ2 were seen in abdominal muscle and heart (P<0.01), low expression levels were seen in leg muscle (P<0.01), longissimus dorsi muscle (P>0.05) and very little expression were detected in liver, spleen, lung and kidney (P>0.05). In addition, high expression levels of MYOZ3 were seen in abdominal muscle, leg muscle, lungs and kidney (P<0.01), low expression levels were found in longissimus dorsi muscle and spleen (P<0.01) and very little expression were detected in heart and liver (P>0.05). Temporal mRNA expression results showed that MYOZ2 and MYOZ3 gene expression varied across four muscle tissues with different ages of the goats. Western blotting further revealed that MYOZ2 and MYOZ3 proteins were only expressed in goat muscle, with notable temporal expression differences in specialized muscle tissues from five development age stages. This work provides the first evidence that MYOZ2 and MYOZ3 genes are expressed abundantly in Tianfu goat muscle tissues from different development age stages, and lay a foundation for understanding the functions of MYOZ2 and MYOZ3 genes in muscle fiber differentiation.

SUBMITTER: Wan L

PROVIDER: S-EPMC3867352 | biostudies-literature | 2013

REPOSITORIES: biostudies-literature

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Molecular cloning and characterization of different expression of MYOZ2 and MYOZ3 in Tianfu goat.

Wan Lu L Ma Jisi J Wang Nianlu N Wang Daihua D Xu Gangyi G

PloS one 20131218 12

The myozenin family of proteins binds calcineurin, which is involved in myocyte differentiation of skeletal muscle. Moreover, gene expression of myozenin is closely related to meat quality. To further understand the functions and effects of myozenin2 (MYOZ2) and myozenin3 (MYOZ3) genes in goat, we cloned them from Tianfu goat longissimus dorsi muscle. Sequence analyses revealed that full-length coding sequence of MYOZ2 consisted of 795 bp and encoded 264 amino acids, and full-length coding seque ...[more]

PMID: 24367523

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Project description:Backgroundβ-defensins have attracted considerable research interest because of their roles in protecting hosts from various pathogens. This study was conducted to investigate the expression profiles of the porcine β-defensin 114 (PBD114) in different breeds and in response to infections. Moreover, the function of PBD114 protein was partially investigated.MethodsSix Tibetan pigs (TP) and six DLY (Duroc×Landrace×Yorkshire) pigs were slaughtered to explore the expression profiles of PBD114 in different breeds and tissues. For infection models, sixteen DLY pigs were divided into two groups and challenged either with sterile saline or E. coli K88. The recombinant protein PBD114 (rPBD114) was obtained by using a heterologous expression system in E. coli.ResultsPBD114 gene was highly expressed in tissues such as the intestine, liver, spleen, and thymus. Interestingly, the expression level of PBD114 gene was higher in the TP pigs than in the DLY pigs (P < 0.05), and was significantly elevated upon E. coli K88 challenge (P < 0.05). The nucleotide sequences of PBD114 from Tibetan and DLY pigs was identical, and both showed a 210-bp open reading frame encoding a 69-amino acid mature peptide. To explaore the function of PBD114 protein, PBD114 gene was successfully expressed in E. coli Origami B (DE3) and the molecular weight of the rPBD114 was estimated by SDS-PAGE to be 25 kDa. The rPBD114 was purified and mass spectrometry verified the protein as PBD114. Importantly, rPBD114 showed antimicrobial activities against E. coli DH5α and E. coli K88, and the minimal inhibitory concentrations (MICs) were 64 and 128 μg/mL, respectively. Hemolytic and cytotoxicity assays showed that rPBD114 did not affect cell viability under physiological concentrations.ConclusionsPBD114 is an infection response gene that is differentially-expressed between different porcine breeds and tissues. The antimicrobial activity of PBD114 protein, against pathogens such as the E. coli K88, suggested that it may serve as a candidate for the substitution of conventionally used antibiotics.

Dataset Information

Molecular cloning and characterization of different expression of MYOZ2 and MYOZ3 in Tianfu goat.

Publications

Molecular cloning and characterization of different expression of MYOZ2 and MYOZ3 in Tianfu goat.

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