Molecular Cloning and Bioinformatics Analysis of TypeⅢSecretion System Effector Protein Va1686 Gene of Vibrio alginolyticus

In this study, Va 1686 gene was cloned from Vibrio alginolyticus . The total length of the gene is 1 164 bp, and it could encode 387 amino acids. The physicochemical properties, protein structure, genetic evolutionary relationship and antigenic characteristics of the effect protein Va1686 of V. alginolyticus HY9901 type Ⅲ secretion system were studied and analyzed by bioinformatics methods and tools. The results showed that Va1686 is a stable hydrophilic and acidic protein without a transmembrane region and a signal peptide, and secondary structure to α-helix. The evolutionary analysis showed that V. alginolyticus HY9901 and V. harveyi were clustered together, which indicated that the genetic relationship between the two species was the closest. Va1686 contains a Fic superfamily conserved domain associated with cell division. Bioinformatics analysis showed that the B-cell preponderant epitopes of Va1686 might be localized in the regions of 48-49, 82-85, 125-126, 150-153, 185-186, 236-237 and so on. The 3D structure model of Va1686 subunit was simulated by SWISS-MODEL software and it was found that the vopS of V. parahaemolyticus was similar and the similarity was 89.46%. In this study, the feasibility of Va1686 as a common antigen of Vibrio was verified from the perspective of bioinformatics, which laid the foundation for the next step in vaccine development.

Cloning and Bioinformatics Analysis of tye A Gene of Vibrio alginolyticus

[Objectives]This study aimed to clone the tye A gene of Vibrio alginolyticus HY9901 strain and analyze its sequence by bioinformatics.[Methods] By referring to the entire genome sequence of Ⅴ.alginolyticus on Gen Bank,specific primers were designed.According to the principle of PCR amplification,the target gene tye A was amplified.By means of bioinformatics,the sequence of tye A was further analyzed,and the phylogenetic tree of tye A genes of Vibrio spp.and the corresponding subunit three-dimensional structure models were constructed.[Results] The length of the tye A gene of Ⅴ.alginolyticus strain HY9901 is 285 bp,and its theoretical molecular weight is 10.98 kD.According to prediction,there is no signal peptide or transmembrane region at the N-terminus of the sequence,and the amino acid sequence contains two casein kinase Ⅱ phosphorylation sites.The results of protein subcellular localization prediction show that the Tye A protein is located in the cell membrane.The protein is unstable and non-hydrophilic.The tertiary structure of Tye A protein of Ⅴ.alginolyticus is similar to that of Yersinia sp.According to prediction,Tye A has a major functional domain Pfam.In terms of secondary structure,alpha helix,random coil,extended strand and beta turn account for 85.11%,7.45%,4.26% and 3.19%,respectively.The homology of Tye A between Ⅴ.alginolyticus and Vibrio parahaemolyticus is up to 83%,so they are classified into one cluster.[Conclusions]This study will help to further understand the regulation mechanism of type Ⅲ secretion system in Ⅴ.alginolyticus.

3D printing of bone tissue engineering scaffolds

Tissue engineering is promising in realizing successful treatments of human body tissue loss that current methods cannot treat well or achieve satisfactory clinical outcomes.In scaffold-based bone tissue engineering,a high performance scaffold underpins the success of a bone tissue engineering strategy and a major direction in the field is to produce bone tissue engineering scaffolds with desirable shape,structural,physical,chemical and biological features for enhanced biological performance and for regenerating complex bone tissues.Three-dimensional(3D)printing can produce customized scaffolds that are highly desirable for bone tissue engineering.The enormous interest in 3D printing and 3D printed objects by the science,engineering and medical communities has led to various developments of the 3D printing technology and wide investigations of 3D printed products in many industries,including biomedical engineering,over the past decade.It is now possible to create novel bone tissue engineering scaffolds with customized shape,architecture,favorable macro-micro structure,wettability,mechanical strength and cellular responses.This article provides a concise review of recent advances in the R&D of 3D printing of bone tissue engineering scaffolds.It also presents our philosophy and research in the designing and fabrication of bone tissue engineering scaffolds through 3D printing.