Department of Physics, Chemistry and Biology (IFM)

In order to study the expression of Dm-dNK, a mouse strain expressing Dm-dNK was constructed. CMV promoter/enhancer was used for the Dm-dNK transgene which was amplified using PCR and cloned into pCDNA3 vector. The transgene was cut from the vector using Bgl II and Dra III restriction enzymes and purified. The purified Dm-dNK transgene was then injected into the female mice using the pronuclear injection technique.

Genotyping was performed by isolating DNA from tail tissues for PCR analysis of the presence of the Dm-dNK gene. The expression of the protein in various tissues was also analysed and the enzymatic activity in both the wild-type mouse (WT C57BL/6) and the Dm-dNK positive mice were determined and compared. Tail samples collected from mice that were approximately 14 days old were cut approximately 0.25 inches from the tip using sterile scissors and the mice were ear marked. DNA was isolated from the tail samples using the DNeasy Blood and Tissue Kit (QIAGEN) and the DNA screened for Dm-dNK gene by PCR using specific primers for the Dm-dNK gene (given below).

Fw : 5’-TAAAGCTTATGGCGGAGGCAGCATCCTGTGC-3’

Rv : 5’-TAGGATCCTTAGTGATGATGATGATGATGTCTGGCGACCCTCTGGCGCTTGCT-3’