cloning of human thap11 gene in lentiviral vector

Milad Esmatparast,1,* Salar khanzad,2

Abstract


Introduction

Thap11, the most recent described member of thap domain family, which is in volved in cell proliferation and described as one of thekey pluripotency gene. despite of very good research projects established to elucidate real role of thap11 in cell, there is still inconsistency among researches about the exact role of thap11 in cells. in this research we are over expressed the thap11 gene in human primary fibroblast cells to find that how this gene could alter the cells behavior during expression

Methods

Human genome was extracted from human blood sample with qiagen dna extraction kit . the thap11 primers designed according to thap11 sequence in ncbi nucleotide database and pcr amplification was done with phusion dna polymerase. the pcr product cloned into pcdh vector by xba1 and ecor1 restriction enzyme digestion followed by purification and ligation steps according to standard protocols and the validity of cloning is proved by colony pcr and dna sequencing

Results

Pcdh letiviral vector containing thap11was constructed and the sequencing analysis proved the sequence of the thap11 gene

Conclusion

Thap11 gene is one of the recently founded pluripotency genes which is very important in stem cells survival and embryo development . so it will be subject to many researches in stem cell and related areas

Keywords

Thap11, ronin, cloni