Part:BBa_K1884003

Hygromycin B

Hygromycin is active against both prokaryotic and eukaryotic cells. It acts by inhibiting polypeptide synthesis. It stabilizes the tRNA-ribosomal acceptor site, thereby inhibiting translocation.

Biology

In the laboratory it is used for the selection and maintenance of prokaryotic and eukaryotic cells that contain the hygromycin resistance gene. The resistance gene is a kinase that inactivates hygromycin B through phosphorylation^[1]. Since the discovery of hygromycin-resistance genes, hygromycin B has become a standard selection antibiotic in gene transfer experiments in many prokaryotic and eukaryotic cells.

HYG is 999bp in length. Fig. 1 shows the DNA sequence of HYG is successfully amplified by PCR from psi-Check2 vector. From this electrophoretogram, we can see the brightness of HYG PCR product is rather high compared with DNA Marker, which indicates that the PCR product of HYG is in a high concerntration

Figure 1 The electrophoretogram of Hygromycin B PCR product.

Usage

In our project, Hygromycin B is for use in Screening monoclone on LB plate. Our team configurated six different concentrations of Hygromycin B solutions in order to figure out the best screening concentration of Paromomycin solution. Due to this experiment, we find out the best screen concentration of Hygromycin B solution is 10ug/ul in LB plate.Fig 2

Figure 2 The colony in LB medium containing Hygromycin B.

Sequence and Features

Assembly Compatibility:

• 10
  COMPATIBLE WITH RFC[10]
• 12
  COMPATIBLE WITH RFC[12]
• 21
  COMPATIBLE WITH RFC[21]
• 23
  COMPATIBLE WITH RFC[23]
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal NgoMIV site found at 455
  Illegal NgoMIV site found at 637
  
• 1000
  INCOMPATIBLE WITH RFC[1000]

  Illegal BsaI.rc site found at 910