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[The construction of NUDC shRNA interference vector with red fluorescence protein]
Xia Liao1, Shi-feng Pang, Qiang Zhang
1Department of Biology of Guangdong Medical College, Zhanjiang, China.
Aim:
Used RFP gene to construct a RNA interference vector for convenience to obtain the good effective hairpins sequence.
Methods:
NUDC and RFP genes were cloned into pDs vector separately, resulting in pDs-NUDC- RFP. as above, human U6 promoter and 9 hairpins sequence of NUDC were cloned into the pDs- NUDC-RFP vector separately.The RNA interfererence vectors target to 9 points of NUDC were constructed. Construct- ed recombinant vectors and then were identified by restrictive digestion and DNA sequencing.293T cells were transfected with the recombinant DNA samples by the liposome complex method, and then fluorescence photographs were taken.
Results:
Enzyme digestion and DNA sequencing showed that the target gene and shRNA fragments were correctly inserted in pDs vector. fluorescence photographs showed that shNUDC-A is the best effective fragment.
Conclusion:
The NUDC gene targeted shRNA and its vector are successfully constructed.
