DPP4 Extracellular Domain Mutation Analysis

The DNA encoding the DPP4 extracellular domain (residues 39–766) was amplified by PCR from the full-length DPP4 gene and subsequently cloned into a modified vector pFastBac1 with six His residues at the C-terminus. Twelve DPP4 mutations, including eight mutations identified from individuals (D65E, P290L, A291P, S323L, T351A, V486M, R492K, and K554Q) of which the detailed information refers to our previous study [25 (link)] and four mutations for confirmatory research (K122D, D243A, P257A, and S664P) were introduced into DPP4 by using phanata master mix DNA polymerase (Vazyme Biotech, catalog number: P515, Nanjing, China), followed by digestion of the template DNA with Dpn I (Thermo Fisher Scientific, catalog number: ER1702, Waltham, MA, USA).

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Li T.T., Peng C., Wang J.Q., Xu Z.J., Su M.B., Li J., Zhu W.L, & Li J.Y. (2021). Distal mutation V486M disrupts the catalytic activity of DPP4 by affecting the flap of the propeller domain. Acta Pharmacologica Sinica, 43(8), 2147-2155.

Publication 2021

Dpn I

Digestion Dna polymerase Dpp4 Gene Mutations Vector

Corresponding Organization :

Other organizations : Shanghai Institute of Materia Medica, National Center for Drug Screening, Chinese Academy of Sciences, Ruijin Hospital, Shanghai Jiao Tong University, ShanghaiTech University

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Variable analysis

independent variables

12 DPP4 mutations including 8 mutations identified from individuals (D65E, P290L, A291P, S323L, T351A, V486M, R492K, and K554Q) and 4 mutations for confirmatory research (K122D, D243A, P257A, and S664P)

dependent variables

Not explicitly mentioned

control variables

The DNA encoding the DPP4 extracellular domain (residues 39–766) was amplified by PCR from the full-length DPP4 gene and subsequently cloned into a modified vector pFastBac1 with six His residues at the C-terminus.

controls

Positive control: Not mentioned.
Negative control: Not mentioned.

Annotations

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