CAJ | 학술논문

目的构建多发性骨髓瘤(MM)黏蛋白MUC1-2VNTR基因的真核表达载体,为研制MM基因疫莳奠定基础.方法以MUC1-2VNTR的编码基因作为目的基因,在其前端加上COZAK序列后,两端分别加上Hind Ⅲ和Xba Ⅰ酶酶切位点,将人工合成的全基因定向克隆入pcDNA3.1/myc-his B中,并通过酶切及测序进行鉴定.结果合成的MUC1-2VNTR基因全长约140 bp,所构建的pcDNA3.1/MUC1-2VNTR/myc-his B重组体经双酶切及测序鉴定后,与预期片断大小相符,并含有完整的阅读框架和目的基因,证明构建成功.结论成功地构建了MM真核表达载体pcDNA3.1/MUC1-2VNTR/myc-his B,为MUC1黏蛋白的功能研究和MM基凶疫苗的研制奠定了相应的实验基础.
목적 구건다발성골수류(MM)점단백MUC1-2VNTR기인적진핵표체재체,위연제MM기인역시전정기출.방법 이MUC1-2VNTR적편마기인작위목적기인,재기전단가상COZAK서렬후,량단분별가상Hind Ⅲ화Xba Ⅰ매매절위점,장인공합성적전기인정향극륭입pcDNA3.1/myc-his B중,병통과매절급측서진행감정.결과 합성적MUC1-2VNTR기인전장약140 bp,소구건적pcDNA3.1/MUC1-2VNTR/myc-his B중조체경쌍매절급측서감정후,여예기편단대소상부,병함유완정적열독광가화목적기인,증명구건성공.결론 성공지구건료MM진핵표체재체pcDNA3.1/MUC1-2VNTR/myc-his B,위MUC1점단백적공능연구화MM기흉역묘적연제전정료상응적실험기출.
Objective To construct multiple myeloma mucin MUC1-2VNTR gene eukaryotic expressing vector,which provided the basic material for further study of multiple myeloma DNA vaccine.Methods MUC1-2VNTR coding gene as target gene,and a KOZAK sequence was inserted before it.Hind Ⅲ and Xba Ⅰ restriction enzyme site were inserted on both ends.Then the whole sequence was synthesized and cloned into pcDNA3.1/myc-his B vector,and the recombinant vector was identified by restriction enzyme digestion and DNA sequencing.Results Synthesized MUC1-2VNTR gene was 140 bp.Restriction enzyme digestion and DNA sequencing confirmed pcDNA3.1/MUC1-2VNTR/myc-his B including the whole exact translation frame region and MUC1-2VNTR gene.Condnsion The pcDNA3.1/MUC1-2VNTR/myc-his B has been successfully constructed,which provides the basic material for further studies of MUC1 mucin function and multiple myloma DNA vaccine.