作为真核核糖体不可或缺的组成部分,核糖体蛋白L23a在蛋白质合成中起着重要作用,折叠和分类。在这项研究中,从小尾寒羊耳缘组织cDNA表达文库中筛选出大小为471bp的核糖体蛋白L23a的cDNA片段,它具有157个氨基酸和17.69kDa的分子量。L23a的核苷酸序列与人类的核苷酸序列具有很高的同源性,鼠标,牛和猪占91.51%,88.32%,96.18%和93.84%,分别。L23a是高度基础的,含有一个合并的45精氨酸,Lys,和His残基,只有14个Asp和Glu残基。用质粒pEGFP-N3-RPL23A转染后,分析了重组L23a蛋白在Ujumqin绵羊成纤维细胞中的表达模式和细胞内分布,转染后24h细胞的细胞质和核仁均有绿色荧光信号,阳性细胞数随时间增加,转染48小时后达到峰值水平。转染效率为22.8%。重组L23a基因在大肠杆菌中的表达模式随诱导温度的不同而不同,电感浓度和诱导时间,当IPTG浓度为0.1mmol/L,诱导温度为37°时,L23a蛋白表达随诱导时间增加。
As an indispensable component of the eukaryotic ribosome, ribosomal protein L23a plays an important role in protein synthesis, folding and sorting. In this study, the cDNA fragment of ribosomal protein L23a with 471 bp in size was screened from the Small Tail Han sheep ear marginal tissue cDNA expression library, it has 157 amino acids and a molecular weight of 17.69 kDa. The nucleotide sequence of L23a shares a high homology with those of human, mouse, cattle and pig of 91.51%, 88.32%, 96.18% and 93.84%, respectively. L23a is highly basic, containing a combined 45 Arg, Lys, and His residues and only 14 Asp and Glu residues. The expression pattern and intra-cellular distribution of recombinant L23a proteins in Ujumqin sheep fibroblast cells were analyzed after transfected with the plasmid pEGFP-N3-RPL23A, there were green fluorescence signals both in the cytoplasm and nucleolus of transfected cells after 24 h, the number of positive cells was increased with time, and they reached the peak level after 48 h of transfection. The transfection efficiency was 22.8%. Expression patterns of recombinant L23a gene in Escherichia coli were different with induction temperature, inductor concentration and induction time, when the IPTG concentration was 0.1 mmol/L and induction temperature was 37°, L23a protein expression was increased with induction time.