Part:BBa_K5023000

Bleomycin Resistance Gene

Resistance to bleomycin is conferred by the product of the Sh ble gene, which was first isolated from Streptoalloteichus hindustanus. The Sh ble gene product binds to the antibiotic in a one-to-one ratio, preventing it from causing DNA cleavage. This resistance gene is utilized as a selectable marker in certain cloning and expression vectors where bleomycin or its analogs, like Zeocin, are used as the antibiotic for selection.

Design

This resistance to bleomycin gene has one rbcs intron. The insertion of an rbcs intron into the bleomycin resistance gene represents an innovative strategy for heterologous expression in Chlamydomonas reinhardtii. The rbcs gene encodes the enzyme ribulose-1,5-bisphosphate carboxylase/oxygenase (RuBisCO), which is essential for the carbon fixation process in photosynthesis. Introns, non-coding DNA sequences, play a crucial role in gene expression regulation and messenger RNA processing.

By inserting an rbcs intron into the bleomycin resistance gene, the aim is to enhance the gene's expression and stability in Chlamydomonas reinhardtii. This strategy may allow the alga to express the bleomycin resistance gene more efficiently, enabling its survival in the presence of the zeocin or bleomicyn antibiotics. Additionally, the intron insertion may help prevent messenger RNA degradation and improve gene translation.

This approach holds significant potential for biotechnology and research in Chlamydomonas reinhardtii. By enhancing the heterologous expression of genes of interest, algal strains with desired characteristics can be developed, ranging from biofuel production to the synthesis of bioactive compounds.

Sequence and Features

Assembly Compatibility:

• 10
  INCOMPATIBLE WITH RFC[10]

  Illegal PstI site found at 227
  
• 12
  INCOMPATIBLE WITH RFC[12]

  Illegal PstI site found at 227
  
• 21
  COMPATIBLE WITH RFC[21]
• 23
  INCOMPATIBLE WITH RFC[23]

  Illegal PstI site found at 227
  
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal PstI site found at 227
  Illegal NgoMIV site found at 608
  Illegal NgoMIV site found at 669
  
• 1000
  COMPATIBLE WITH RFC[1000]

References

GATIGNOL, A.; DURAND, H.; TIRABY, G. Bleomycin resistance conferred by a drug-binding protein. FEBS Letters, v. 230, n. 1-2, p. 171–175, 28 mar. 1988.