ABSTRACT
Objective: To clone the full-length cDNA of the ATP/ADP transporter protein (AATP) genes in Panax ginseng to provide resources and some knowledge necessary for the further gene function study. Methods: The mRNA sequence of the AATP genes in other plants were downloaded on the website of NCBI and used to perform local Blast alignment in the transcriptome of Jilin ginseng from 14 tissues. The AATP gene in Panax ginseng was cloned by PCR, and analyzed using bioinformatics software and online resources. The expression pattern of PgAATP1 gene in 14 tissues of Panax ginseng was analyzed using the expression profile of transcriptome and its expression level under methyl jasmonate was deceted by quantitative real-time PCR. Results: A full-length cDNA sequence was successfully cloned from Panax ginseng and named as PgAATP1, which was 1866 bp in length and encoded 621 amino acids. The relative molecular mass of PgAATP1 protein calculated was 67 897.23, and the isoelecric point calculated was 9.58. It was found that the protein was similar to the plastid AATP in other species. The expression of this gene was high in all tissues but higherin fruit flesh and leaf blade, and the expression of PgAATP1 gene was up-regulated by methyl jasmonate. Conclusion: We have obtained the full-length of PgAATP1 gene. This gene expressed higher in tissues of vigorous starch synthesis and responsing to methyl jasmonate.
ABSTRACT
Recent studies have shown that diet can affect the body's immunity. Roughage of dairy cows consists of a variety of plant materials which make different contributions to health. This study investigated the effect of different roughages on the immunity of dairy cows. Serum, peripheral blood mononuclear cells (PBMCs), and milk samples were collected from 20 multiparous mid-lactation cows fed mixed forage (MF)- or corn straw (CS)-based diets. Expression profile analysis was used to detect the differentially expressed genes (DEGs) from PBMCs. The results showed that milk protein in the MF group increased to 3.22 g/100 ml, while that of the CS group milk was 2.96 g/100 ml; by RNA sequencing, it was found that 1615 genes were differentially expressed between the CS group and the MF group among the 24 027 analyzed probes. Gene ontology (GO) and pathway analysis of DEGs suggested that these genes (especially genes coding cytokines, chemokine and its receptors) are involved in the immune response. Results were confirmed at the protein level via detecting the levels of interleukin-2 (IL-2), IL-6, IL-10, IL-12, leptin (LEP), interferon-γ (IFN-γ), transforming growth factor-β1 (TGF-β1), and tumor necrosis factor-α (TNF-α) in peripheral blood by enzyme-linked immunosorbent assay (ELISA) and radioimmunoassay analysis. Our data supported the conclusions that the protein content in milk of the MF group was higher than that of the CS group, the CS-based diets induced more release of cytokines than the MF-based diets in dairy cows' PBMCs, and milk protein content may be affected by cytokines.