Extraction of high-quality genomic DNA from wild boar tissue is of great significance for the study of its genetic information, population quantity, and distribution. In this work, a hydrophobic magnetic deep eutectic solvent (HMDES) was synthesized. Based on the HMDES, a HMDES-based vortexed extraction method was developed for extracting genomic DNA from wild boar muscle tissue. Compared to the traditional high salt-extraction method, this approach is faster and embraces higher DNA extraction efficiency. Different pretreatment methods before extraction were evaluated. Single-factor experiments were used to optimize the extraction condition, for example, extraction time, temperature, and HMDES volume. After extraction, the DNA can be quickly and easily recovered from the HMDES phase, and the HMDES can be reused. This work provides a simple and environmental-friendly extraction method for DNA extraction from wild boar tissue.
Glutathione S -transferases (GSTs) play a key role in detoxification of xenobiotics in organisms. However, their other functions, especially response to the natural toxin cantharidin produced by beetles in the Meloidae and Oedemeridae families, are less known. We obtained GST cDNAs from three sources: Cydia pomonella ( CpGSTd1 ), Sus scrofa ( SsGSTα1 ), and Triticum aestivum ( TaGSTf3 ). The predicted molecular mass is 24.19, 25.28 and 24.49 kDa, respectively. These proteins contain typical N-terminal and C-terminal domains. Recombinant GSTs were heterologously expressed in Escherichia coli as soluble fusion proteins. Their optimal activities are exhibited at pH 7.0–7.5 at 30 °C. Activity of CpGSTd1 is strongly inhibited by cantharidin and cantharidic acid, but is only slightly suppressed by the demethylated analog of cantharidin and cantharidic acid. Enzymatic assays revealed that cantharidin has no effect on SsGSTα1 activity, while it significantly stimulates TaGSTf3 activity, with an EC 50 value of 0.3852 mM. Activities of these proteins are potently inhibited by the known GST competitive inhibitor: S -hexylglutathione (GTX). Our results suggest that these GSTs from different sources share similar structural and biochemical characteristics. Our results also suggest that CpGSTd1 might act as a binding protein with cantharidin and its analogs.
Iron deficiency is an important public health concern worldwide. Intake of iron-fortified foods has been widely used to treat iron deficiency anemia (IDA). In this study, a novel food for iron supplementation was designed: pig (Sus scrofa domestica) skin peptide-iron (PSP-Fe) chelates. Structural characterization demonstrated that acidic amino acids (aspartic acid, glutamic acid) and aromatic amino acids (phenylalanine, tryptophan, and tyrosine) in PSP were involved in the chelation reaction, with the carboxyl group and amino group provided the major iron binding sites. In addition, iron significantly altered the microscopic morphology of PSP. IDA rats were established and different doses of iron supplements were gavaged for 21 days to evaluate the effectiveness of PSP in treating IDA. The medium dose of PSP-Fe restored hemoglobin (HGB), red blood cell (RBC), hematocrit (HCT), mean corpuscular hemoglobin concentration (MCHC), serum ferritin (SF), serum iron (SI), hepcidin, total iron binding capacity (TIBC) and transferrin saturation (TSAT) to normal levels. PSP-Fe also ameliorated the abnormal changes in heart coefficients, lungs coefficients, liver coefficients and spleen coefficients caused by IDA. PSP-Fe further restored iron storage in the liver and villous damage in the colon of rats compared to FeSO4. 16S rRNA results suggest that the 10 microbial markers in the Model group may impede iron absorption and HGB synthesis of host through biosynthesis of siderophore group nonribosomal peptides, vitamin B6 metabolism, lipoic acid metabolism, ascorbate metabolism and tryptophan metabolism. At the end, the safety of PSP-Fe was preliminarily affirmed by toxicity evaluation in vitro and in vivo. These findings suggest that PSP-Fe has potential as a novel functional food for treating IDA.
危险品化学品经营许可证（带存储）