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The aim of the present study was based on the IL-2 gene signal peptide coding sequence , different modified optimization , and compare its ability to mediate secretory expression of human endostatin adenovirus vector , screened an efficient mediated endostatin gene expression secretion of an adenovirus vector , lay the foundation for the development of long-term , stable and low side effects of endostatin adenovirus - anti-tumor gene therapy . Endo Method 1 , respectively, for wild-type IL-2 signal peptide , optimized IL-2 signal peptide , the Kozak sequence of the wild-type IL-2 signal peptide , four signal peptide boot Kozak sequence optimized IL-2 signal peptide gene (IwE , IME , Kiwe , KIME ) ; 2 , were constructed for the four recombinant plasmids were co-transfected pDC315 ( io ) - IWE , pDC315 ( io ) - IME pDC315 - Kiwe ( io ) , pDC315 ( io ) - KIME ; 3 , the use of HI - IQ system restructuring four recombinant adenovirus rAd - IWE the rAd - IME , the RAD - Kiwe , rAd - KIME , amplification, purification and titer was determined ; 4 , four recombinant adenovirus vector analysis and comparison of expression and expression levels in NCI-H460 cells . The results obtained four signal peptide the Endo gene boot ; 2, build - IWE for transfection of four recombinant plasmid pDC315 ( io ) , pDC315 ( io ) -IME , pDC315 ( io ) - Kiwe pDC315 ( io)-KImE; 3, to obtain the rAd-IwE, rAd-ImE, rAd-KIwE, rAd-KImE four recombinant adenovirus titer was determined order of 3.9 × 10 8,3.7 × 10 8,3.1 × 10 - 8,3.5 × 10 8 ; analysis to get Ad - the IME , Ad - KIME two efficient mediated secretion of recombinant adenovirus vector expressing Endo , the highest expression than Ad - IWE 3 times , 4 times . Conclusion We have obtained more efficient -mediated secretion of endostatin expression of recombinant adenovirus vector .
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