Part:BBa_K1898400

CRYAB, crystallin alpha B

This gene codes for CRAYB, also known as crystallin alpha B, a crystallin protein exists in the lens and also other tissues and organs.

Usage and Biology

Alpha crystallin is the most abundant protein in the lens. Out of the two types of alpha crystallin proteins, alpha B is less restricted to the lens and has a 1:3 ratio to alpha A [1]. It has been found that crystallin alpha B affects lens development and has chaperone activity to maintain normal protein interaction[2].

In our project, we made CRYAB because we wanted to monitor crystallin protein's response to oxidative stress and to test our prevention and treatment for cataracts.

[1]Genes and mapped phenotypes. (n.d.). from https://www.ncbi.nlm.nih.gov/gene/1410

Sequence and Features

Assembly Compatibility:

• 10
  COMPATIBLE WITH RFC[10]
• 12
  COMPATIBLE WITH RFC[12]
• 21
  INCOMPATIBLE WITH RFC[21]

  Illegal BamHI site found at 26
  Illegal BamHI site found at 368
  
• 23
  COMPATIBLE WITH RFC[23]
• 25
  COMPATIBLE WITH RFC[25]
• 1000
  INCOMPATIBLE WITH RFC[1000]

  Illegal BsaI site found at 453
  Illegal SapI.rc site found at 313
  

Gel Picture

We designed primers to move CRYAB cDNA into iGEM BioBrick. PCR was set up and the gel picture is shown below. The expected band size is ~1kb, and the bands at the expected size are boxed in red.

Sequencing

We sent CRYAB out to sequencing after it's cloned into iGEM backbone. The sequencing file is annotated as follows:

• four restriction sites are highlighted in red
• CRYAB is highlighted in orange

This confirms that the sequence is correct (with no stop codon) and is moved into the BioBrick.