遗传学报
遺傳學報
유전학보
ACTA GENETICA SINICA
2000年
11期
1006-1011
,共6页
曾君祉%周志勇%吴有强%刘志萍%王文星%黄华梁%蔡祝南%于嘉林
曾君祉%週誌勇%吳有彊%劉誌萍%王文星%黃華樑%蔡祝南%于嘉林
증군지%주지용%오유강%류지평%왕문성%황화량%채축남%우가림
BNYVV%杂交瘤%单链抗体%E. coli%表达
BNYVV%雜交瘤%單鏈抗體%E. coli%錶達
BNYVV%잡교류%단련항체%E. coli%표체
BNYVV%hybridoma%single-chain Fv antibody%E. coli%expression
用PCR方法以分泌抗甜菜坏死黄脉病毒（BNYVV）单克隆抗体的杂交瘤细胞的基因组为模板，扩增了编码BNYVV单抗的重链可变区（VH）基因。测序表明，该VH序列属于小鼠II（A）亚类，全长为360bp，编码120个氨基酸。将其和先前克隆的轻链基因分别插入到一个含有连接VH和VL基因的连接序列的质粒之中，构建成单链抗体（scFv）基因的表达载体pTCscFv。将质粒在大肠杆菌中表达，ELISA法检测出表达产物有与BNYVV抗原结合的活性。
用PCR方法以分泌抗甜菜壞死黃脈病毒（BNYVV）單剋隆抗體的雜交瘤細胞的基因組為模闆，擴增瞭編碼BNYVV單抗的重鏈可變區（VH）基因。測序錶明，該VH序列屬于小鼠II（A）亞類，全長為360bp，編碼120箇氨基痠。將其和先前剋隆的輕鏈基因分彆插入到一箇含有連接VH和VL基因的連接序列的質粒之中，構建成單鏈抗體（scFv）基因的錶達載體pTCscFv。將質粒在大腸桿菌中錶達，ELISA法檢測齣錶達產物有與BNYVV抗原結閤的活性。
용PCR방법이분비항첨채배사황맥병독（BNYVV）단극륭항체적잡교류세포적기인조위모판，확증료편마BNYVV단항적중련가변구（VH）기인。측서표명，해VH서렬속우소서II（A）아류，전장위360bp，편마120개안기산。장기화선전극륭적경련기인분별삽입도일개함유련접VH화VL기인적련접서렬적질립지중，구건성단련항체（scFv）기인적표체재체pTCscFv。장질립재대장간균중표체，ELISA법검측출표체산물유여BNYVV항원결합적활성。
The heavy chains variable region gene (VH) of monoclonal antibody against beet necrotic yellow vein virus (BNYVV) was amplified from total DNA extracted from anti-BNYVV hybridoma cells by PCR. Sequencing showed that the VH belongs to mouse subgroup II(A) and contains 360bp, which code one hundred and twenty amino acids. The VH and VL genes were inserted into a plasmid which contains a linker sequence for constructing scFv gene. The new vector named pTC scFv. The scFv was produced in Escherichia coli and appeared binding activity with BNYVV antigen by ELISA method.
