Studies on the Physiological Function of Lymphocytes and the Polymorphism of Major Histocompatibility Complex in Japanese Quails

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1.Studies on the lymphocyte antigens of Japanese quail : (1) As hybridomer of 5 clones derived from thymocyte : QT-1-QT-5 ; and 5 clones derived from bursacyte : QB-1-QB-5.(2) Imminohistochemical analysis on the immuno organ were carried out by the monoclonal anitibodies. QT-1^+, -4^+ and 5^+ cells only exist incortex of thymus.QB-1^+, -2^+ and -3^+ cells exist in medulla or bursa of Fabricius and in B cell locus of PALS of spleen.(3) Lymphocyte antigens were analyzed by flowcytometory. QT-1^+, -4^+ and 5^+ cells only exist thymus and their frequencies were 70 - 80%. The frequencies of QB-1^+, -2^+ and -3^+ cells were 90% in brusacytes.2.Analysis of major histocompatibility complex (MHC) class I gene from Japanese quail : (1) RFLPs patten of genomic DNA from Japanese quail which were analyzed using BF10 in chicken MHC class I cDNA as a probe showed polymorphism.(2) Nine positive clones : QF41,63,76,95,103,111,113,128 ; corresponding to the MHC class I gene of Japanese quail were isolated by screening of liver cDNA library with BF10 and QF41 as arobe. Based on comparison among sequences of QF clones, MHC class I of Japanese quail classified into 5 loci.(3) PCR analysis of alpha1-alpha2 region using genomic DNA as a template with QF63 and QF41 specific primers were revealed different sizes of fragment by 609bp in PCR product of QF63 vs 434bp in QF41 product. QF63 has 181bp intron between alpha1 and alpha2 regions, but QF41 have not intron between alpha1 and alpha2 regions.(4) PCR products using the QF63 specific primer were digested with the 2 kinds of restriction endnuclease : Ban II and EcoT14 I.Combination of PCR-RFLP patterns using 2 kinds of restriction endnuclease classified 9 haplotypes QF63 product, but only 5 haplotypes ovserved in this experiment.