Description
1) ASD Background
Autism Spectrum Disorder (ASD) is a neuro-developmental disorder characterized by social communication disorders,
narrow interests and activities, and repetitive behaviors. Generally, ASD starts in infancy and mainly manifests in
different extents of social skills and communication skills, accompanied by certain intellectual and behavioral
disorders.
According to recent data, in 2014, the disease rate of ASD in US infants is 2.24%. However, in 2016, the disease rate increased to 2.76%, which means that the disease rate increased from year to year. Nowadays, the number of Chinese people with ASD peaks at 14 million.
According to recent data, in 2014, the disease rate of ASD in US infants is 2.24%. However, in 2016, the disease rate increased to 2.76%, which means that the disease rate increased from year to year. Nowadays, the number of Chinese people with ASD peaks at 14 million.
The reasons contributed to ASD are complicated. Heredity, Abnormal brain structure and function, biological
mechanism, adverse factors of pregnancy, and environmental factors are common factors that cause ASD to occur. There
are many ways to diagnose ASD, but none of them are convenient and cheap, and there’s still no targeted medicine for
ASD. The biggest problem in curing ASD is age. The higher the age, the higher the difficulty in therapy.
We find a great need in the diagnosis portion and medicine portion of ASD, so we think about a solution to help people.
In further research, we find out that patients usually have a high value in glutamate, and tryptophan and these two elements become our signs. SLC7A5 is a protein that can ease the symptoms. From these information, we think manipulating a kit and a kind of probiotic beverage will be a good solution.
We find a great need in the diagnosis portion and medicine portion of ASD, so we think about a solution to help people.
In further research, we find out that patients usually have a high value in glutamate, and tryptophan and these two elements become our signs. SLC7A5 is a protein that can ease the symptoms. From these information, we think manipulating a kit and a kind of probiotic beverage will be a good solution.
2) Experiment
Our purposes are to detect signs of ASD (Kit) and ease the symptoms (Probiotic beverage)
To finish our tasks, we need to construct 3 plasmids.
1. pET28a +SLC7A5 can ease the symptoms
2. pTrc99k+tnaC+amilGFP can determine the value of tryptophan
3. pET28a+inaK-gldh can help determine the value of glutamate
The first plasmid is a therapeutic plasmid, and the rest of the two are detection plasmids.
To construct the plasmids, we let the company synthesize the pET28a+inaK-gldh plasmids. We amplified tnaC and amilGFP DNA fragments and insert them into the pTrc99k vector, and amplified SLC7A5 and inserted it into the pET28a vector.
Next, we extracted the plasmids and transformed them into the competent bacteria E. coli BL21(DE3). We verified the protein expression level by SDS-PAGE and purified the proteins.
Finally, we verified if our biosensor could be used to detect ASD by measuring the fluorescence intensity of L-tryptophan. We also measured the activity of inaK-gldh.
To finish our tasks, we need to construct 3 plasmids.
1. pET28a +SLC7A5 can ease the symptoms
2. pTrc99k+tnaC+amilGFP can determine the value of tryptophan
3. pET28a+inaK-gldh can help determine the value of glutamate
The first plasmid is a therapeutic plasmid, and the rest of the two are detection plasmids.
To construct the plasmids, we let the company synthesize the pET28a+inaK-gldh plasmids. We amplified tnaC and amilGFP DNA fragments and insert them into the pTrc99k vector, and amplified SLC7A5 and inserted it into the pET28a vector.
Next, we extracted the plasmids and transformed them into the competent bacteria E. coli BL21(DE3). We verified the protein expression level by SDS-PAGE and purified the proteins.
Finally, we verified if our biosensor could be used to detect ASD by measuring the fluorescence intensity of L-tryptophan. We also measured the activity of inaK-gldh.
3) Expected Result
1. Successfully construct pET28a +SLC7A5, pTrc99k+tnaC+amilGFP, and pET28a+inaK-gldh plasmids.
2. Expressed and purified inaK-gldh and SLC7A5 protein.
3. The inaK-gldh enzyme could use L-glutamate as a substrate.
4. We could measure the fluorescence intensity of the pTrc99k+tnaC+amilGFP plasmid contain strain when adding L-tryptophan.
2. Expressed and purified inaK-gldh and SLC7A5 protein.
3. The inaK-gldh enzyme could use L-glutamate as a substrate.
4. We could measure the fluorescence intensity of the pTrc99k+tnaC+amilGFP plasmid contain strain when adding L-tryptophan.
4) Reference
1. Deth R, Muratore C, Benzecry J, et al. How environmental and genetic factors combine to cause autism: a
redox/methylation hypothesis. Neurotoxicology, 2008, 29: 190–201
2. Tomova A, Husarova V, Lakatosova S, et al. Gastrointestinal microbiota in children with autism in Slovakia. Physiol Behav, 2015, 138: 179-87
3. Dora C. Tărlungeanu, et al. Impaired Amino Acid Transport at the Blood Brain Barrier Is a Cause of Autism Spectrum Disorder.cell.2016
4. Chen L, Shi X-J, Liu H, Mao X, Gui L-N, Wang H, et al. Oxidative stress marker aberrations in children with autism spectrum disorder: a systematic review and meta-analysis of 87 studies (N = 9109). Transl Psychiatry. (2021) 11:15.
2. Tomova A, Husarova V, Lakatosova S, et al. Gastrointestinal microbiota in children with autism in Slovakia. Physiol Behav, 2015, 138: 179-87
3. Dora C. Tărlungeanu, et al. Impaired Amino Acid Transport at the Blood Brain Barrier Is a Cause of Autism Spectrum Disorder.cell.2016
4. Chen L, Shi X-J, Liu H, Mao X, Gui L-N, Wang H, et al. Oxidative stress marker aberrations in children with autism spectrum disorder: a systematic review and meta-analysis of 87 studies (N = 9109). Transl Psychiatry. (2021) 11:15.