Difference between revisions of "Part:BBa K3863006"
Line 14: Line 14:
 
<partinfo>BBa_K3863006 SequenceAndFeatures</partinfo>
 
<partinfo>BBa_K3863006 SequenceAndFeatures</partinfo>
  
−
<h2>Reference</h2>
+
==Team PuiChing_Macau 2021: Break down monosaccharides==
 +
===Validation of construct expression using RT-qPCR===
 +
https://2021.igem.org/wiki/images/7/7e/T--PuiChing_Macau--1aa.jpg
 +
<p>1a) Amylase (BBa_K3863006) qPCR</p>
 +
 
 +
https://2021.igem.org/wiki/images/9/92/T--PuiChing_Macau--1b.jpg
 +
<p>1b) Glucoamylase (BBa_K3863006) qPCR</p>
 +
 
 +
<p>
 +
To future confirm that our constructs have the expression level, we have performed RT-qPCR
 +
The RT-qPCR results show that we have successfully assembled target genes K3863006 to the vector (pETDuet).
 +
</p>
 +
 
 +
==Reference==
 
<p>[1]Masayoshi Sakaguchi,corresponding author1 Yudai Matsushima,1 Toshiyuki Nankumo,1 Junichi Seino,1 Satoshi Miyakawa,1 Shotaro Honda,1 Yasusato Sugahara,1 Fumitaka Oyama,1 and Masao Kawakita1,2, (2014) (Glucoamylase of Caulobacter crescentus CB15: cloning and expression in Escherichia coli and functional identification)</p>
 
<p>[1]Masayoshi Sakaguchi,corresponding author1 Yudai Matsushima,1 Toshiyuki Nankumo,1 Junichi Seino,1 Satoshi Miyakawa,1 Shotaro Honda,1 Yasusato Sugahara,1 Fumitaka Oyama,1 and Masao Kawakita1,2, (2014) (Glucoamylase of Caulobacter crescentus CB15: cloning and expression in Escherichia coli and functional identification)</p>
  

Revision as of 15:30, 21 October 2021


a-amylase-HA-glucoamylase-his

This is a composite part designed to break down starch in food waste into monosaccharides. Amylase: E. coli periplasmic α-amylase gene malS. The protein sequence in this part is based on K523001 from Edinburgh 2011. Glucoamylase: Caulobacter crescentus CB15, caulobacter vibrioides gene for glucoamylase(GenBank: AB813000)

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 1287
    Illegal NotI site found at 2146
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal XhoI site found at 3884
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 2441
    Illegal NgoMIV site found at 2633
    Illegal NgoMIV site found at 2637
    Illegal NgoMIV site found at 3275
    Illegal NgoMIV site found at 3649
    Illegal NgoMIV site found at 3803
    Illegal NgoMIV site found at 3824
    Illegal AgeI site found at 423
    Illegal AgeI site found at 1588
    Illegal AgeI site found at 2393
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 3026

Team PuiChing_Macau 2021: Break down monosaccharides

Validation of construct expression using RT-qPCR

T--PuiChing_Macau--1aa.jpg

1a) Amylase (BBa_K3863006) qPCR

T--PuiChing_Macau--1b.jpg

1b) Glucoamylase (BBa_K3863006) qPCR

To future confirm that our constructs have the expression level, we have performed RT-qPCR The RT-qPCR results show that we have successfully assembled target genes K3863006 to the vector (pETDuet).

Reference

[1]Masayoshi Sakaguchi,corresponding author1 Yudai Matsushima,1 Toshiyuki Nankumo,1 Junichi Seino,1 Satoshi Miyakawa,1 Shotaro Honda,1 Yasusato Sugahara,1 Fumitaka Oyama,1 and Masao Kawakita1,2, (2014) (Glucoamylase of Caulobacter crescentus CB15: cloning and expression in Escherichia coli and functional identification)