Part:BBa_K1613007
A composite part that can detect some toxins.
Introduction
cyp1a2 can oxidize the substances that it can metabolism.
Cytochrome P450 1A2 is a member of the cytochrome P450 mixed-function oxidase system. And it is involved in the metabolism of xenobiotics in the body. Function The cytochrome P450 proteins are monooxygenases which catalyze many reactions involved in drug metabolism and synthesis of cholesterol, steroids and other lipids. CYP1A2 is able to metabolize some PAHs to carcinogenic intermediates. Other xenobiotic substrates for this enzyme include aflatoxin B1, and acetaminophen.
CYP1A2 Substrates
many antidepressants amitriptyline clomipramine, some atypical antipsychotics clozapine olanzapine, ropivacaine, theophylline, zolmitriptan, melatonin, tamoxifen, erlotinib, cyclobenzaprine, estradiol, fluvoxaminemexiletine, naproxen, ondansetron, phenacetin, paracetamol, propranolol, riluzole, tacrine, tizanidine, verapamil, warfarin
Design
When aflatoxin B1 goes into the e.coli, CYP1A2 will oxidize the Aflatoxin B1, and then the aflatoxin B1 will turn into Aflatoxin oxidation AFBO. The Aflatoxin oxidation will cause DNA damage, making DNA become single-strand DNA(ssDNA), which can activate protein RecA.
E.coli has a sos response system, and there is a repressor LexA which inhibits the next reaction, so
the red fluorescent gene won’t express. But when the activated protein RecA comes and hydrolysis the repressor LexA, the next reaction can work, so the fluorescent gene will express. Therefore, we can know that there is Aflatoxin B1 in the oil we test. The eleven wells from left to right:Well one to seven are K1613003(CYP1A2+B0015); the others are K1613005.
We try three different RBS(B0030,B0034,B0032) to increase the protein expression.
Sources
NCBI, GeneBank:NM000761
References
[1]Veronica S. Mary a, Ana Valdehita b, Jose M. Navas b, Hector R. Rubinstein a, Maria L. Fernandez-Cruz. Effects of aflatoxin B1, fumonisin B1 and their mixture on the aryl hydrocarbon receptor and cytochrome P450 1A induction [2]Yan-Ping Du Hui-Lian Zhu. The possible mechanism of Aflatoxin B1 Hepatocarcinogenesis and preventive effects of phytochemicals [3]Food hygiene standards - Food in aflatoxin limits (human) 82 1.4.4 Health Department Announcement fresh word No. 8189322 [4]EU animal and plant inspection and quarantine of food hygiene regulations cum Profile [5]Veronica S. Mary a, Ana Valdehita b, Jose M. Navas b, Hector R. Rubinstein a,*Maria L. Fernandez-Cruz b. Effects of aflatoxin B1, fumonisin B1 and their mixture on the aryl hydrocarbon receptor and cytochrome P450 1A induction [6]ZHENG Lili, ZHU Yujing, SHAO Caimei, ZHANG Yong The Relationship between Cytochrome CYP3A4 in Vivo and the Toxicity of Aflatoxin B1 in Feedstuffs [7]Nejc Paulič2,†,Adrijana Leonardi1, Vesna Hodnik2,Gregor Anderluh2,3, Zdravko Podlesek2, Darja Žgur-Bertok2,Igor Križaj1,4,5 Matej Butala2. Structural insight into LexA–RecA* interaction Lidija Kovačič1
Sequence and Features
- 10INCOMPATIBLE WITH RFC[10]Illegal prefix found in sequence at 64
Illegal suffix found in sequence at 1653 - 12INCOMPATIBLE WITH RFC[12]Illegal EcoRI site found at 64
Illegal NheI site found at 7
Illegal NheI site found at 30
Illegal SpeI site found at 1654
Illegal PstI site found at 1668
Illegal NotI site found at 70
Illegal NotI site found at 1661 - 21INCOMPATIBLE WITH RFC[21]Illegal EcoRI site found at 64
Illegal BglII site found at 1483 - 23INCOMPATIBLE WITH RFC[23]Illegal prefix found in sequence at 64
Illegal suffix found in sequence at 1654 - 25INCOMPATIBLE WITH RFC[25]Illegal prefix found in sequence at 64
Illegal XbaI site found at 79
Illegal SpeI site found at 1654
Illegal PstI site found at 1668
Illegal NgoMIV site found at 403
Illegal NgoMIV site found at 490
Illegal AgeI site found at 473 - 1000INCOMPATIBLE WITH RFC[1000]Illegal BsaI.rc site found at 1203
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