Study Myocilin (MYCO) Gene Exon 3 Polymorphisms Of A Genealogy With Primary Open Angle Glaucoma (POAG) In Xining,

Objective: To analyze and study Myocilin (MYCO) genes single nucleotide polymorphisms of a genealogy with primary open angle glaucoma (POAG) in Xining, detect the mutation of gene exon 3.Method: 1.To check systemically the ocular pressure, vision, fundus, visual field, angle and do other ophthalmologic examinations for the patient of primary open angle glaucoma, his family and healthy people.2. To extract the whole Genome DNA: use DNA isolation kit to extract peripheral blood of the glaucoma patient, his family and healthy people. To detect the extracted DNA through spectrophotometer and agarose gel electrophoresis.3.Amplify the area of MYCO gene exon3: using the whole Genome DNA as the Template, amplify 3 pairs of specific primers which compounded in accordance with MYCO gene exon 3 by polymerase chain reaction. 4. Evaluation of PCR products: evaluate PCR products by agarose gel electrophoresis. 5. Purification of PCR products: using QIAquick Gel Kit to purify the ampliative products.6.To directly analyze the gene sequencing and single nucleotide polymorphisms of purified products.Result: After the examination of genealogy of primary open-angle glaucoma, we have found all the MYCO gene of glaucoma and suspected patients can be detected point mutation of Pro370Leu in exon 3 area, that is the 370th amino acid of MYCO protein will change from Pro to Leu, and no one was found in contrasted ten healthy cases.Conclusion: One mutant site of was detected in exon 3 area of MYCO of the glaucoma genealogy patient and caused the 370th amino acid of MYCO protein changed from Pro to Leu; the change of this single nucleotide polymorphism caused structural and functional change of MYCO protein. It might be the important factor which caused the genealogical primary open-angle glaucoma. Extract peripheral blood DNA, amplify exon 3 of MYCO gene with PCR reaction, determine disease development through single nucleotide polymorphism and analyze and study its mutation condition. This method is convenient, sensitive and reliable and can be one of the basic methods of screening glaucoma patient.