Wet lab

1. New parts to the registry. We submitted two of our parts to the iGEM registry which are useful for genetic engineering: BBa_K4786000 and BBa_K4786001. BBa_K4786000 is the promoter of Vibrio natriegens PilA gene, which can be used for the detection of the expression level of the PilA gene. For future iGEM teams who want to work with Vibrio natriegens, and who are interested in its pilus, this part could be of help. And we also tested the suppression efficiency of two sgRNA target sites within this part.BBa_K4786001 contains a spdCas9 gene controlled by a trc promoter. This part is useful for producing dCas9 protein in bacteria. We demostrated that this composite part could function in Vibrio natriegens.

1. 向iGEM库中添加了新的部件。我们提交了两个实用基因工程部件：BBa_K4786000和BBa_K4786001。BBa_K4786000是需钠弧菌PilA基因的启动子，可用于检测PilA表达水平的报告系统中，以检测调控系统的作用。BBa_K4786001包含了由trc 启动子控制的dCas9基因，能够在细菌中表达dCas9蛋白。

2. An ideal CRISPRi system and the corresponding reporter system. The aim of this system is to exclude the impact of V. Natriegens’ own pili. To achieve the ideal effect, we designed and tested several different versions and finally achieved the proper inhibition of the expression of V. Natriegens’ own pili, and meanwhile enable the test of the effect of CRISPRi through our sfGFP-based reporter system. These two systems both provide basis for relative future researches.

1. 向iGEM库中添加了新的部件。我们提交了两个实用基因工程部件：BBa_K4786000和BBa_K4786001。BBa_K4786000是需钠弧菌PilA基因的启动子，可用于检测PilA表达水平的报告系统中，以检测调控系统的作用。BBa_K4786001包含了由trc 启动子控制的dCas9基因，能够在细菌中表达dCas9蛋白。

2. An ideal CRISPRi system and the corresponding reporter system. The aim of this system is to exclude the impact of V. Natriegens’ own pili. To achieve the ideal effect, we designed and tested several different versions and finally achieved the proper inhibition of the expression of V. Natriegens’ own pili, and meanwhile enable the test of the effect of CRISPRi through our sfGFP-based reporter system. These two systems both provide basis for relative future researches.

3. 得到了一个生产导电菌毛的新底盘微生物。在该项目中我们发展了一个产生导电菌毛的的需钠弧菌菌株。作为目前已知的生长速度最快的菌种，需钠弧菌能够满足大规模工业生产的要求，并且其作为新兴的底盘微生物具有显著优势。通过多轮的实验及故障排除，我们获得了能够产生金属还原地杆菌导电菌毛的需钠弧菌菌株，并将我们的实验方法呈现在“experiment"部分，供未来参考。

Model

In the model, we constructed a molecular dynamics model for the type IV pili of G.metallireducdens. Based on this model, we used algorithms to optimize two template functions that we set with the aim of obtaining conductive pili with higher conductivity. Our main contributions in this section are as follows:

在Model中，我们构建了金属还原地杆菌的Ⅳ型的分子动力学模型，基于该模型我们使用算法分别对于两个我们设定的模板函数分别进行优化，希望能够得到具有更高电导率的导电菌毛。我们在该板块的主要贡献如下：

1. Established the first molecular dynamics model for the type IV pili (e-pili) of G.metallireducdens. The PDB file for this protein can be downloaded via the following link

1.建立了第一个金属还原地杆菌的Ⅳ型菌毛（导电菌毛）的分子动力学模型。该蛋白的PDB文件可以点击此链接下载

G.m e-pili.pdb

2. Established a mathematical model for searching and optimizing the internal electron transfer chain within the pili based on specified objective functions. Subsequent teams can use our model to optimize other types of e-pili. Additionally, the process of building this model can provide possible insights for the performance optimization of structural proteins in other team projects. The code we used in this process can be downloaded via the following link

2.建立了根据指定目标函数搜索并优化菌毛内部电子传递链的数学模型，后续团队可以使用我们的模型来优化其他种类的导电菌毛。除此之外，该模型的建立过程有助于为其他团队项目中的结构蛋白的性能优化提供可能的思路。我们在此过程中使用到的代码可以点击此链接下载

APSO.py Training.py

3. Designed a biofuel cell screening system that can be used to select potentially higher e-pili. With this system, we can implement semi-rational directed evolution of the e-pili protein.

设计了一套可以用于筛选可能具有的更高电导率菌毛的生物燃料电池筛选系统。配合该系统我们可以实现对于该导电菌毛蛋白的半理性定向进化。

Hardware

In the Hardware section, in order to put our system into use, we designed and built the supporting hardware to realize the real-time detection and data analysis of the conductivity of E-PIli at different concentrations of the disease markers to be measured. Our main contributions in this area are as follows.

1. PCB design for resistance detection with WiFi module.
2. Structure and casing design and manufacturing files of Plinker
3. Structure and casing design and manufacturing files of E-Pili Cartridge
4. Experimentation and documentation of biodegradable electrode materials

Integrated Human Practices

We have developed a framework for Integrated Human Practices, covering planning, execution and demonstration. The framework is reuseable for other teams and can be seen as guidance. We have named this framework the Plink Framework.