Part:BBa_K4887006

HygR

It is a hygromycin resistance gene.

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 AgeI site found at 683
  
• 1000
  COMPATIBLE WITH RFC[1000]

6.5 Results:

(1) Selection of postively transformed calli with hygromycin

When enough of sweet potato embryogenic callus ware obtained, they were infected with the A. tumefaciens transformants containing the vector for CRISPR-knockout expression by co-culture (Fig. 1A). After selection with hygromycin, positive transformed calli were obtained (Fig. 1B).

Fig. 11 Embryogenic callus co-culture with A. tumefaciens transformants (A) and selection of postively transformed calli with hygromycin (B)

(2) PCR detection for gene HygR.

Genome DNA of the transgenic lines was extracted from leaves of these two regenerated seedlings. PCR detection on the genomes was performed by using primers designed based on the HygR protein gene. As the result, the transgenic lines 23216004 and 23216005 were further validated (Fig. 2). The sequences of the primers were as bellow:

• HygR-F: 5’-Atgaaaaagcctgaactcac-3’
• HygR-R: 5’-ctatttctttgccctcggac-3’

Fig. 2 Detection results of PCR for gene HygR.