Expression of Mutant EctodysplasinA Gene in Hypohidrotic Ectodermal Dysplasia

• Published in: International journal of human genetics Vol. 22; no. 3; p. 128
• Main Authors: He, Fangqi, Wang, Hongfeng, Lu, Shouyi, Gao, Qingping, Guo, Feng, Lei, Ye, Zeng, Tingwen
• Format: Journal Article
• Language: English
• Published: Delhi Kamla-Raj Enterprises 01.09.2022
• Subjects: Amino acids; Dysplasia; E coli; Ectodysplasin; EDA protein; Enzymes; Gene expression; Mutants; Mutation; Plasmids; Point mutation; Protein expression; Proteins
• ISSN: 0972-3757
• DOI: 10.31901/24566330.2022/22.03.814

T To investigate the protein expression changes of c.878T>G mutant and wild-type Ectodysplasin-A (EDA) gene on prokaryotic vectors from hypohidrotic ectodermal dysplasia (HED) families. The c.878T>G mutant and wild-type EDA genes were amplified, screened, and double-enzyme digested to obtain the target fragments. The recombinant plasmids were constructed and introduced into Escherichia coli. Lactosidase (IPTG) induces the expression of the target gene. The EDA protein expression content was determined by Western blot and enzyme-linked immunosorbent method. On the prokaryotic expression vector, the c.878T>G mutant and wild-type EDA protein expression increased with the increase of induction time, and the mutant protein expression was lower than the wild-type protein expression at the same point (P<0.05). In conclusion, the c.878T>G gene mutation in this family caused one amino acid of the EDA protein to become another amino acid, and the c.878T>G mutation site can lead to a significant decrease in the expression of EDA protein on the prokaryotic expression vector, is highly pathogenic and may be the cause of the HED family.