CAJ | 학술논문

用PCR方法获得编码人组织型纤溶酶原激活物（tissure-type plasminogen activator ，t-PA）C端P区肽段的DNA，并经测序证实，长810 bp，含有全部的t-PA的丝氨酸蛋白酶编码序列.将这段序列克隆到表达载体pQE30中转化大肠杆菌JM109菌株,经SDS-PAGE检测：转化菌株中表达出分子量约29×103的蛋白，用纤维平板法测定激活纤溶活性，结果表明表达的t-PA蛋白C端P区具有很强的激活纤溶蛋白酶原的活性.证实t-PA蛋白C端P区的酶结构区的功能是独立的,其生物活性不受其它区域影响．
용PCR방법획득편마인조직형섬용매원격활물（tissure-type plasminogen activator ，t-PA）C단P구태단적DNA，병경측서증실，장810 bp，함유전부적t-PA적사안산단백매편마서렬.장저단서렬극륭도표체재체pQE30중전화대장간균JM109균주,경SDS-PAGE검측：전화균주중표체출분자량약29×103적단백，용섬유평판법측정격활섬용활성，결과표명표체적t-PA단백C단P구구유흔강적격활섬용단백매원적활성.증실t-PA단백C단P구적매결구구적공능시독립적,기생물활성불수기타구역영향．
The DNA fragment encoding t-PA P region protein was obtained by PCR,using plasmid pTAhtpa(F) harborong the full length t-PA cDNA as template. This P region fragmemt was cloned into pQE30 and transformed to JM109 strain.P region proteins fused to 6 His peptides at its N terminus with molecular weight of 29×103 were expressed in JM109 induced by IPTG.The expressed protein were purified by Ni-NTA-agarose and were studied its activity by fibrin plate activity assay(FPAA).The results show that the function of t-PA P region is indepent for other regions of t-PA. and it has significant activity to stimulate plasminogen.