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Yangzhou University

11 ARTICLES PUBLISHED IN JoVE

Biochemistry

Biosynthesis of a Flavonol from a Flavanone by Establishing a One-pot Bienzymatic Cascade

Zhiping Zhang¹, Shuhang Fan¹, Zhiqiang Chen¹, Yanzhi He¹, Mengfei Huang¹, Li Ding¹, Yanyan Zhang⁵, Lin Chen¹, Xinyue Zhang^1,2,3,4

¹College of Bioscience and Biotechnology, Yangzhou University, ²Key Laboratory of Prevention and Control of Biological Hazard Factors (Animal Origin) for Agrifood Safety and Quality, Ministry of Agriculture of China, Yangzhou University, ³Joint International Research Laboratory of Agriculture & Agri-Product Safety, Yangzhou University, ⁴Jiangsu Co-innovation Center for Prevention and Control of Important Animal Infectious Diseases and Zoonoses, ⁵The Testing Center, Yangzhou University

The derivation of a flavonol is crucial for its application in healthcare and the food industry. Here, we provide a detailed protocol for the biosynthesis of a flavonol from a flavanone and discuss the crucial steps and its advantages over other approaches.

Biology

Measuring Crop Motility and Food Passaging in Drosophila

Jiadong Cai^*1,2, Junmeng Xi^*1,2, Youheng Wei^1,2

¹Joint International Research Laboratory of Agriculture and Agri-Product Safety, the Ministry of Education of China, Yangzhou University, ²College of Bioscience and Biotechnology, Yangzhou University

The goal of this protocol is to measure crop contraction and quantify food distribution in the Drosophila gut.

Immunology and Infection

Use of Fimbrial Rod for F18ab Fimbriae⁺ STEC Colonization to Host Cells

Mingxu Zhou^*1,2, Qiangde Duan^*1,3, Yang Yang^1,3, Guoqiang Zhu^1,3

¹College of Veterinary Medicine, Yangzhou University, ²Institute of Veterinary Immunology & Engineering, Jiangsu Academy of Agricultural Sciences, ³Jiangsu Co-innovation Center for Prevention and Control of Important Animal Infectious Diseases and Zoonoses

Here we present a protocol to study the function of fimbriae in bacterial colonization.

Immunology and Infection

A Non-Coding Small RNA MicC Contributes to Virulence in Outer Membrane Proteins in Salmonella Enteritidis

Xia Meng^*1,2, Weiwei Cui^*1, Xianchen Meng¹, Jianye Wang^1,2, Jinqiu Wang³, Guoqiang Zhu^1,2

¹College of Veterinary Medicine, Yangzhou University, ²Jiangsu Co-innovation Center for Prevention and Control of Important Animal Infectious Diseases and Zoonoses, ³Department of Animal Husbandry and Veterinary Medicine, Beijing Agricultural Vocational College

An λ-Red-mediated recombination system was used to create a deletion mutant of a small non-coding RNA micC.

Biology

A Simple Dry Sectioning Method for Obtaining Whole-Seed-Sized Resin Section and Its Applications

Jiajing Qiu^*1,2, Yinhui Ren^*1,2, Lingxiao Zhao^1,2, Biao Zhang^1,2, Cunxu Wei^1,2

¹Key Laboratory of Crop Genetics and Physiology of Jiangsu Province/Jiangsu Key Laboratory of Crop Genomics and Molecular Breeding, Yangzhou University, ²Co-Innovation Center for Modern Production Technology of Grain Crops of Jiangsu Province/Joint International Research Laboratory of Agriculture & Agri-Product Safety of the Ministry of Education, Yangzhou University

This technique allows for the fast and simple preparation of whole-seed-sized resin section for the observation and analysis of cells, starch granules, and protein bodies in different regions of the seed.

Immunology and Infection

Production of Monoclonal Antibodies Targeting Aminopeptidase N in the Porcine Intestinal Mucosal Epithelium

Pengpeng Xia^1,2,3, Siqi Lian^1,2,3, Yunping Wu^1,2,3, Li Yan^1,2,3

¹College of Veterinary Medicine (Institute of comparative medicine), Yangzhou University, ²Jiangsu Co-innovation Center for Prevention and Control of Important Animal Infectious Diseases and Zoonoses, ³Joint International Research Laboratory of Agriculture and Agri-Product Safety of Ministry of Education of China, Yangzhou University

The recombinant antibody protein expressed in pIRES2-ZSGreen1-rAbs-APN-CHO cells and monoclonal antibodies produced using traditional hybridoma technology can recognize and bind to the porcine aminopeptidase N (APN) protein.

Biology

A Non-Degradative Extraction Method for Molecular Structure Characterization of Bacterial Glycogen Particles

Jun-Jiao Wang^*1, Meng-Meng Wang^*2,3, You-Wei He⁴, Zhang-Wen Ma^2,3, Zi-Yi Wang^5,6,7, Rui Qiao^2,3, Xin-Le Tan^5,6, Qing-Hua Liu⁸, Liang Wang^1,2

¹Department of Bioinformatics, School of Medical Informatics and Engineering, Xuzhou Medical University, ²Jiangsu Key Laboratory of New Drug Research and Clinical Pharmacy, School of Pharmacy, Xuzhou Medical University, ³Department of Pharmaceutical Analysis, School of Pharmacy, Xuzhou Medical University, ⁴Department of Biological Sciences, School of Life Sciences, Xuzhou Medical University, ⁵School of Chemistry and Molecular Biosciences, The University of Queensland, ⁶Centre for Nutrition and Food Sciences, Queensland Alliance for Agriculture and Food Innovation, The University of Queensland, ⁷Joint International Research Laboratory of Agriculture and Agri-Product Safety, College of Agriculture, Yangzhou University, ⁸State Key Laboratory of Quality Research in Chinese Medicines, Macau University of Science and Technology

Bacterial glycogen structure is greatly impacted by extraction methods which may result in molecular degradation and/or biased sampling. It is essential to develop methods to minimize these problems. Here, four extraction methods have been compared using size distribution and chain length distribution as key criteria for minimizing extraction artifacts.

Biochemistry

The Extraction of Liver Glycogen Molecules for Glycogen Structure Determination

Ziyi Wang^1,2, Qinghua Liu^3,4, Liang Wang^3,5, Robert G. Gilbert^1,2,6, Mitchell A. Sullivan⁷

¹School of Chemistry and Molecular Biosciences, The University of Queensland, ²Centre for Nutrition and Food Sciences, Queensland Alliance for Agriculture and Food Innovation, The University of Queensland, ³Jiangsu Provincial Key Laboratory of New Drug Research and Clinical Pharmacy, School of Pharmacy, Xuzhou Medical University, ⁴Department of Pharmaceutical Analysis, School of Pharmacy, Xuzhou Medical University, ⁵Department of Bioinformatics, School of Medical Informatics and Engineering, Xuzhou Medical University, ⁶Joint International Research Laboratory of Agriculture and Agri-Product Safety, College of Agriculture, Yangzhou University, ⁷Glycation and Diabetes Group, Mater Research Institute-The University of Queensland, Translational Research Institute

An optimal sucrose concentration was determined for the extraction of liver glycogen using sucrose density gradient centrifugation. The addition of a 10 min boiling step to inhibit glycogen-degrading enzymes proved beneficial.

Developmental Biology

In Ovo Intravascular Injection in Chicken Embryos

Kai Jin^1,2, Jing Zhou^1,2, Gaoyuan Wu^1,2, Ziyi Lian^1,2, Zongyi Zhao^1,2, Shujian Zhou^1,2, Chen Chen^1,2, Hongyan Sun^1,2, Yingjie Niu^1,2, Qishenng Zuo^1,2, Yani Zhang^1,2, Jiuzhou Song³, Guohong Chen^1,2, Bichun Li^1,2,4

¹Joint International Research Laboratory of Agriculture and Agri-Product Safety of Ministry of Education of China, Yangzhou University, ²Key Laboratory of Animal Breeding Reproduction and Molecular Design for Jiangsu Province, College of Animal Science and Technology, Yangzhou University, ³Animal & Avian Sciences, University of Maryland, College Park, ⁴College of Biotechnology, Jiangsu University of Science and Technology

The overall goal of this paper is to describe how to perform in ovo intracellular injection of exogenous materials into chicken embryos. This approach is very useful to study the developmental biology of chicken embryos.

Biology

AirID-Based Proximity Labeling for Protein-Protein Interaction in Plants

Ahmad Zada^*1, Imran Khan^*1, Min Zhang¹, Yuting Cheng¹, Xueyun Hu¹

¹College of Bioscience and Biotechnology, Yangzhou University

Here, we present a step-by-step protocol for performing the proximity labeling (PL) experiment in cucumber (Cucumis sativus L.) using AT4G18020 (APRR2)-AirID protein as a model. The method describes the construction of a vector, the transformation of a construct through agroinfiltration, biotin infiltration, protein extraction, and purification of biotin-labeled proteins through affinity purification technique.

Immunology and Infection

A Mouse Ear Model for Allergic Contact Dermatitis Evaluation

Haifeng Shao^1,2,3,4, Jialin Deng^2,3,4, Yanqing Liu^1,2,3,4, Niuniu Yang^1,2,3,4

¹Department of Traditional Chinese Medicine, The Affiliated Hospital of Yangzhou University, Yangzhou University, ²Department of Traditional Chinese and Western Medicine, Medical College, Yangzhou University, ³The Key Laboratory of Syndrome Differentiation and Treatment of Gastric Cancer of the State Administration of Traditional Chinese Medicine, Yangzhou University, ⁴Jiangsu Key Laboratory of Integrated Traditional Chinese and Western Medicine for Prevention and Treatment of Senile Diseases, Yangzhou University

Here, we describe the methods for inducing allergic contact dermatitis in mouse ears by 1-fluoro-2,4-dinitrobenzene (DNFB) and how to evaluate the severity of allergic contact dermatitis.