Difference between revisions of "Part:BBa K4093001"

Revision as of 10:28, 21 October 2021

PUS

PUS

Profile

Name: PUS

Base Pairs: 294bp

Origin: lactic acid bacteria

Properties: Secreted protein

Usage and Biology

BBa_K4093001 is a coding sequence of lactic acid bacteria, which can help protein to be secreted outside the cell.

Construct design

Figure 1. Design diagram.

Our project is to design an edible "drink" for monogastric animals, mainly poultry through biosynthesis technology. The core product will be a probiotic that can produce xylanase. In this project, we designed to construct a plasmid expressing xylanase gene, then we transformed them into Escherichia coli and Lactobacillus reuteri for further performance analysis. Compared to traditional feed, our drink contains the probiotics that could help poultry digest xylan, thus increasing feed efficiency. In this way, not only can the time and economic cost of feeding be saved, but also the gastrointestinal tract of the animal can be protected and thus the disease rate can be reduced.

Experimental approach

Figure 2. electrophoregram of enzyme-digested products of pSIP403 and PUS Channel 1~2: pSIP403, 6kbp, correct Channel 3~4: PUS, 300bp, correct.

Proof of function

Plasmid pSIP403-PUS-xynA transformed into L. reuteri to for secretory expression of xynA in L. reuteri. As shown in Table 1, significant xylanase activity was detected from broken supernatant by DNS method. This indicates that our PUS protein can help xynA protein to be secreted outside the cell.

Table 1. Xylanase activity (OD540 nm) was detected from broken supernatant by DNA method.

Future plan

We aim at developing a probiotic (Lactobacillus) containing xylanase to produce feed additives and poultry beverages, which are believed to help poultry digest xylan and enhance their health. Maybe it is possible for us to produce a novel “mixed feed” that combines other feed additives to enhance more poultry and ruminants’ digestion. After in-depth research, we obtained a feasible plan of “mixed feed” and the details of this plan can be seen at our wiki page partnership.

References

1.Beasley S S,Takala T M， Reunanen J， Apajalahti J， Saris P E.2004.Characterization and electrotransformation of Lactobacillus crispatus isolated from chicken crop and intestine. Poult Science，83(1)：45-48;

2.De Vos W M.1999.Gene expression systems for lactic acid bacteria. Current Opinion in Microbiology，2(3)：289-295;

3.Silversides F G， Scott T A， Korver D R，Afsharmanesh M，Hruby M.2006. A study on the interaction of xylanase and phytase enzymes in wheat-based diets fed to commercial white and brown egg laying hens.Poultry Science，85(2)：297-305;

4.崔罗生，祝茂生，徐顺清，朱辉，梁运祥，张忠明.2009.黑曲霉木聚糖酶基因(xyn A) 在大肠杆菌中的表达及酶学分析.华中农业大学学报，28(1)：48-53；

==== 5.李慧.2010. 蛋白酶和木聚糖酶对肉鸡生长性能，消化机能及血液指标的影响.[研究生学位论文].杨凌：西北农林科技大学； 6.https://wenku.baidu.com/view/bb1a6d76590216fc700abb68a98271fe910eafb9.html ====

Sequence and Features