( 3 ) at post - translation level plant mutual sequence of starting translation aaca and eukaryotic secretory signal peptide sequence was added to 5 ' - flanking region of t - pa gene and kdel ( sequence located to endoplastic reticulum ) to 3 ' - flanking region by pcr amplication and plant expression vector pbemt was constructed ( 3 )翻译后水平通过pcr扩增的方式在t - pa基因5端添加了真核分泌信号肽序列和植物翻译起始共有序列aaca ,在3端添加了内质网定位序列kdel ,构建了植物表达载体pbemt 。
The new synthesized protein was led to endoplastic reticulum cavity by eukaryotic secretory signal peptide sequence and then anchored to innerwall of endoplastic reticulum by kdel sequence , which interdicted the process of protein entering golgi body and cytoplasm , and then avoided heterogeneous glycosylation modification of foreign protein and prolonged the disappearance of half life of protein in organism . 2 真核分泌信号肽序列可以引导新合成的蛋白质进入内质网腔, kdel序列将进入内质网腔的蛋白质锚定在内质网内壁上,从而阻断了蛋白质进入高尔基体和细胞质的过程,进而避免了外源蛋白质的异源糖基化修饰,延长了蛋白质在生物体内的半衰期。
The length of this phytase gene is1506bp interrupted once by an intron of 102bp in the 5 " part of the gene , this intron contains donor sequence - gtatgc , lariat sequence - gctgac and acceptor sequence - cag which are typically conserved sequence of the intron of fungal phytase gene . this gene encodes a peptide of 467amino acid residues with molecular weight of 51 . 37kda , containing 13 potential n - glycosylation sites and a signal peptide sequence made up of 19 amino acid residues at n teminal of the peptide 核苷酸序列分析表明, pcr扩增产物中包含有完整的phya基因,该基因全长1506bp ,其中包含一段长102bp的内含子,该内含子具有真菌植酸酶基因内含子的特征保守序列: donor序列? gtatgc , lariat序列? gctgac及acceptor序列? cag 。该基因编码467个氨基酸,理论分子量为51 . 37kda ,其上有13个潜在的n -糖基化位点, n端19个氨基酸为信号肽序列,植酸酶活性位点序列( crvtfaqvlsrhgaryptdskgk )位于氨基酸序列的+ 71 + 93 。
In comparison with genbank data , the homologies of the nucleotide sequence and amino acid sequence were as following : hc was 98 . 4 % and 100 % ; ha was 97 . 2 % and 99 . 3 % respectively . 4 . two recombinant prokaryotic expression vector were constructed which has complete open reading occlusing initation codon , leader signal peptide sequence and termination codon 序列分析表明,所克隆获得的基因与genbank中已经登录的核苷酸和氨基酸的同源性分别为: hc98 . 4和100 , ha97 . 2和99 . 3 ,证明本试验虫株与国外报道的同源性很高。
Therefore , blys , its receptor or related antagonists may find medical utility in the treatment of b cell disorders associated with autoimmunity , neoplasia , or immunodeficiency syndromes . in this study , epo signal peptide sequence and hsblys gene were linked by soe method . the fusion product was cloned into eukaryotic plasmids . pcdna3 , pcdna3 . 1 , pefneo , respectively . meanwhile , the epo signal peptide sequence was mutated so as to form a restriction enzyme cut site : bin i . thus the recombinant plasmid can be used as secreting plasmid expressing other gene 本实验通过3 ’端互补,进行引物延伸合成epo信号肽序列:信号肽和hsblys基因采用重叠延伸拼接法形成融合基因;融合基因分别插入pcdna3 . 0 、 pcdna3 . 1 、 pefneo真核载体:引物延伸合成信号肽时,利用亮氨酸同义密码,将信号肽基因的倒数第二个密码突变,在重组载体上的信号肽序列之后,形成bln酶切位点,使三种载体成为分泌表达载体。