Strauss M, Bartsch F O, Stollwerk J, Trstenjak M, Böhning A, Gassen H G, Machleidt W, Turk V
Institut für Biochemie der Technischen Hochschule Darmstadt.
Biol Chem Hoppe Seyler. 1988 May;369 Suppl:209-18.
A DNA containing the coding sequence for the human cysteine proteinase inhibitor protein cystatin C has been obtained by enzymatic ligation of chemically synthesized deoxyoligonucleotides, using the Khorana ligation method. The 375 bp synthetic gene carries signals for the translation initiation and termination and was expressed in E. coli as a beta-galactosidase fusion protein as well as a secreted protein under the control of the E. coli alkaline phosphatase signal sequence. The secreted hybrid protein was shown to have similar biological properties as the authentic protein isolated from human plasma.
通过使用霍拉纳连接法对化学合成的脱氧寡核苷酸进行酶促连接，已获得了包含人半胱氨酸蛋白酶抑制剂胱抑素C编码序列的DNA。这个375bp的合成基因带有翻译起始和终止信号，并在大肠杆菌中作为β-半乳糖苷酶融合蛋白以及在大肠杆菌碱性磷酸酶信号序列控制下的分泌蛋白表达。所分泌的杂合蛋白显示出与从人血浆中分离出的天然蛋白相似的生物学特性。
