CAJ | 학술논문

为了研究猪 Siglec-10（pSiglec-10）的分子生物学特征，利用 Ensemble 网站提供的预测序列（ENSSSCT00000032460）设计一对特异性引物，采用 RT-PCR 技术，克隆 pSiglec-10分子的全长 CDS 区，经过测序比较鉴定正确后，对该分子的基本特征进行分析；构建截短 pSiglec-10的原核表达载体，另将该分子的 CDS 区全长克隆至真核表达载体 pcDNA3．1-3′FLAG，将重组质粒转染至 PK-15和 Marc-145细胞中分析其表达定位特征。结果克隆的 pSiglec-10分子的全长 CDS 为2127 bp，编码709个氨基酸；pSiglec-10基因序列与猴的同源性最高，同源性达73．6％，与鼠的同源性为66．9％；与人的同源性为62．5％，氨基酸序列的同源性分别为58．9％、69．4％和65．3％；pSiglec-10 C 端的 ITIM 区域和磷酸化位点高度保守；pSiglec-10分子具有较好的抗原特性和亲水性；该分子主要表达定位在细胞浆中，在 Marc-145细胞中呈不连续的点状分布于细胞浆，细胞核周围荧光强度较强。pSiglec-10分子全长 CDS 区的克隆和分析将对该分子的功能研究提供一定的理论依据。
위료연구저 Siglec-10（pSiglec-10）적분자생물학특정，이용 Ensemble 망참제공적예측서렬（ENSSSCT00000032460）설계일대특이성인물，채용 RT-PCR 기술，극륭 pSiglec-10분자적전장 CDS 구，경과측서비교감정정학후，대해분자적기본특정진행분석；구건절단 pSiglec-10적원핵표체재체，령장해분자적 CDS 구전장극륭지진핵표체재체 pcDNA3．1-3′FLAG，장중조질립전염지 PK-15화 Marc-145세포중분석기표체정위특정。결과극륭적 pSiglec-10분자적전장 CDS 위2127 bp，편마709개안기산；pSiglec-10기인서렬여후적동원성최고，동원성체73．6％，여서적동원성위66．9％；여인적동원성위62．5％，안기산서렬적동원성분별위58．9％、69．4％화65．3％；pSiglec-10 C 단적 ITIM 구역화린산화위점고도보수；pSiglec-10분자구유교호적항원특성화친수성；해분자주요표체정위재세포장중，재 Marc-145세포중정불련속적점상분포우세포장，세포핵주위형광강도교강。pSiglec-10분자전장 CDS 구적극륭화분석장대해분자적공능연구제공일정적이론의거。
In order to explore the functions of pSiglec-10,here we designed a pair of primers according to the predict sequence from Ensemble database to clone whole CDS region of pSiglec-10,then the basic infor-mation of pSiglec-10 was analyzed.A prokaryotic expression vector of truncated pSiglec-10 was recon-structed,the targeting clone was screened correctly and cloned into expression vector pcDNA3.1-3′FLAG, the reconstructing plasmid was transfected into cell PK-15 and Marc-145 to analyze the location of pSiglec-10 in cells.As a result,a full length CDS clone of pSiglec-10 with 2 127 bp was screened correctly,and it has a similarity with human,macaque and mouse in 62.5%,73.6%,66.9%,respectively.The consistency of the amino acid sequence of pSiglec-10 was compared with other three species in 58.9%,69.4%,65.3%, respectively.This molecule expressed in cytoplasm and distributed in discontinuous punctate collection in cell Marc-145,and showed a better antigenicity and hydrophilicity.This research would be a reference to explore the functions of pSiglec-10.