α-GAL
1.Lectin BS-I-B4 is used to mimic the human xenogeneic natural antibody since the capacity of binding Gal-α1,3-Gal. So it can be used to select Gal-α1,3-Gal mimikers.
2.a-D-galactosidase ( a-Gal, R. C. 3. 2. 1. 22 ) i s an exo-glycosidaso, widely present in the bio-resources.
3.Adding 0.1% Tween-80, Triton-100, or EDTA to the culture markedly improved GAL activity.
4.In the study, the p P gal-Control vector, the pTAL-ERE-luc . vector the pCNX2-hERa/pCNX2-hERp. vector were transfected into HEK293 cell line.
6.Result: Transformants can grow on medium of SD/-Leu/-Trp/-His/-Ade/X-α-Gal and the colonies are blue. α? β- galactosidase assays show that the activity of α?
7.Diploid yeast was plated on synthetic dropout nutrient medium (SD/-Trp-Leu-His-Ade) containing x-α-gal.
8.Results Parallel results were obtained from these pig tissues stained against human sera and BSI- B4.There was no significant dif- ference both in the antigens recognized by sera of different blood types or BSI- B4and in the distribution ofα- Gal.
9.CONCLUSION: The stable expression of human α-galactosidase in NIH 3T3 cell strongly inhibits the expression of Gal epitopes, resulting in abrupt reduction in xenorejection induced by human serum.
10.The cDNA of u -Gal gene from C. canephora is 99.27% homologous to the published C. arabica sequence,while 8bp of DNA sequence or 4 AA of protein sequence were changed.
