Author: Tan, Zhaoli; Gao, Lihua; Wang, Yan; Yin, Huihui; Xi, Yongyi; Wu, Xiaojie; Shao, Yong; Qiu, Weiyi; Du, Peng; Shen, Wenlong; Fu, Ling; Jia, Ru; Zhao, Chuanhua; Zhang, Yun; Zhao, Zhihu; Sun, Zhiwei; Chen, Hongxing; Hu, Xianwen; Xu, Jianming; Wang, Youliang
Title: PRSS contributes to cetuximab resistance in colorectal cancer Document date: 2020_1_1
ID: tymoeyoo_70
Snippet: Human SPINK1 coding regions were PCR-amplified from a human mammary retroviral complementary DNA library and cloned into the prokaryotic expression vector pET22b. The primers used were as follows: forward, 5′-GAATTCTAATGGACTCCCTGGGAAGA-GAGGCC-3′; reverse, 5′-CTCGAGGCAAGGCCCAGATTTTTGAA-3′. The fusion protein SPINK1 was expressed in Escherichia coli and purified by nickel affinity and size exclusion chromatography. Isopropyl-ï¢-d-thiogalac.....
Document: Human SPINK1 coding regions were PCR-amplified from a human mammary retroviral complementary DNA library and cloned into the prokaryotic expression vector pET22b. The primers used were as follows: forward, 5′-GAATTCTAATGGACTCCCTGGGAAGA-GAGGCC-3′; reverse, 5′-CTCGAGGCAAGGCCCAGATTTTTGAA-3′. The fusion protein SPINK1 was expressed in Escherichia coli and purified by nickel affinity and size exclusion chromatography. Isopropyl-ï¢-d-thiogalactopyranoside (0.2 mM/liter) induction at 16°C for 16 hours was estimated to be an optimal expression strategy. The expression of the fusion proteins was detected by Western blotting and Coomassie Brilliant Blue G-250 staining.
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