黃顙魚基礎(chǔ)免疫的研究.pdf

1、華中農(nóng)業(yè)大學碩士學位論文黃顙魚基礎(chǔ)免疫的研究姓名：黃艷青申請學位級別：碩士專業(yè)：水產(chǎn)養(yǎng)殖指導(dǎo)教師：陳孝煊王桂堂20030501黃艷青黃顙魚基礎(chǔ)免疫的研究AbstractThreedifferentisolatingmethodswereappliedtofindthebestwaytopurlfyserumlgMfromthebullhesdcatfish，Pelteobagrus聲lvidracointhepresentstudyTh

2、epurityandmolecularweightoftheserumIgMweredetermined，andthentherabbitpolyclonalanfiserawerepreparedwiththepurifiedIgMInaddition，themicrostructureandujlrastructureof出eperipheralbloodcellsandthethymusofthebullheadcatfishwe

3、reobservedanddescribedAlltheseresultsweuldbenefitforthefurtherstudyoftheimmunologyofthebullheadcatfishInthepurificationoftheserulrltgMfromtheimmunisedandnonimmunisedt7shthreekindsofaffmitychromatographiccolumnwereusedThe

4、yweremannanbindingprotein(MBP)affirtitychromatographiccolumn，TGaffinitychromatographiccolumnandimmunoaffmitychromatographiccolumnAmongthemMBP蕊血“chromatogra[)hiccolumnandimmuneammtychromatographiccolumnwereeffectivetoobta

5、lnabundantIgMwi也highpufftybutt11elatterneededlongin2muisedtimeTGaffinitychromatographiccolumnwasalsoaneffectivemethodforpurlfyingbutwithIowquantityoflgMoutcomeBySDSpolyacrylamidegelelectrophoresis(SDSPAGE)，themolecularwe

6、ightoftheIgMHchainandlgMLchainwasdeterminedthevaluebeing724kDand299kD、respectivelyRabbitantibullheadcatfishIgMpolyclonalantiserawereproducedbyimmunizingrabbitwi出thepurifiedIgM，Tiireofthepolyclonalantiseraweremeasuredbyus

7、ingsandwichELISATotalserumproteinandtheconcentrationoflgMofthenormalseraofbullheadcatfishweredeterminedbyusingtheCooma；sicbluedyebindingmethodofBradfordandsandwichELlSA，thevaluesbeingt7，57mg／mtandl，16mg／ml，respectively11

8、1epercentageofIgMinthetotalserumproteinwas660％。OnthestainedperipheralbloodSmearsofthebullhcadcatfislLseventypesofbloodcellswererecognizedTheywereerythrocyte，thrombocyte，lymphocyte，monocyte、neutrophil，basophilandeosinophi

9、lMoreoverthegranulocytescouldbeclassifiedintotwotypes，ThethymusofthebutlheadcatfishlocatedatthebackofthegillchamberinsidetheopercularepitheliumandoutsidetheepioticopharyngoclaviculariesAwelldevelopedthymusofthefishcallbe

10、dividedintothreepans：theouterzonemiddleZoneandinnerzoneTheouterzonewasmadeupofretieularcells，colmnnarcellsandmucouscells；whilethemiddiezonewascharacterizedwithlargernumberofdeeplystainedandcompactedlymphocyteInthemiddlea

11、ndinnerzonestherewereseveralHassallscorpuscles，whichweqremadeupoftwoormoreendothelialceltssurroundingartemptychamberoramastcellSomesinailholesonthethymusmembranec0111dbeobserved，whichmightberesponsiblefortheantigeningest

12、ionThethymusoflyearoldfishwassupposedbyconnectivetrabccula’andthatof2yearold丘shshowedapparentlydegeneratingnlethymusofthe3yearoldfishjustremainedaspeciallydegeneratedstructureKeywords：Pelteobagrusfulvidraco；affinitychrom