秦川牛和中國(guó)荷斯坦牛FSHβ基因SSCP多態(tài)性與雙胎性狀的關(guān)系研究.pdf

1、摘要1秦川牛和中國(guó)荷斯坦牛FSHp基因SSCP多態(tài)性與雙胎性狀的關(guān)系研究摘要本實(shí)驗(yàn)以秦川牛和中國(guó)荷斯坦牛的雙胎性狀為研究對(duì)象，采用SSCP分子標(biāo)記方法對(duì)FSHp基因的多態(tài)性進(jìn)行了分析，并結(jié)合測(cè)序?qū)ζ浠虻?’端、第3外顯子和3端進(jìn)行了序列分析，旨在探索牛雙胎性狀的分子標(biāo)記的適宜方法，為今后高繁殖力牛的選育提供一定的理論依據(jù)。研究結(jié)果如下:LFSHp基因5端多態(tài)性及序列分析秦川牛在FSH日基因5端的三對(duì)引物(FSH(3P1FSHpP2FS

2、HpP3)擴(kuò)增片段均有多態(tài)，中國(guó)荷斯坦牛除了引物FSH(3PI所擴(kuò)增片段沒(méi)有檢測(cè)到多態(tài)外，在其兩對(duì)引物擴(kuò)增片段中均發(fā)現(xiàn)存在多態(tài)。(1)對(duì)于引物FSHRPI擴(kuò)增片段，在秦川牛上檢測(cè)到AABB和AB型三種基因型，群體多態(tài)信息含量(PIC)達(dá)到了中度多態(tài)〔0.5PIC0.25)，秦川牛群體中多態(tài)性高，遺傳變異較大，而在中國(guó)荷斯坦牛中只檢測(cè)到了1種基因型，該位點(diǎn)未發(fā)現(xiàn)多態(tài)。測(cè)序結(jié)果顯示，秦川牛中AA基因型個(gè)體在65bp插入了一個(gè)G堿基BB基因型

3、個(gè)體在73bp281bp和284bp堿基處發(fā)生了突變，73bp處插入了一個(gè)T堿基，281bp處TG284by處GA.(2)對(duì)于引物FSHpP2擴(kuò)增片段，在秦川牛和中國(guó)荷斯坦牛上均檢測(cè)到了多態(tài)性，均出現(xiàn)了正常型和突變型個(gè)體，秦川牛正常型個(gè)體在642bp處GA突變型個(gè)體在642bp處GA6906p處CG。在中國(guó)荷斯坦牛中正常型個(gè)體和突變型個(gè)體在642bp處的突變與秦川牛一致，均發(fā)生GA的突變，另外突變型個(gè)體在424bp處還插入了堿基Ca(3

4、)I物FSHOP3擴(kuò)增片段在被檢測(cè)的秦川牛和中國(guó)荷斯坦牛群體中均表現(xiàn)出了多態(tài)。測(cè)序結(jié)果顯示，秦川牛和中國(guó)荷斯坦牛個(gè)體在1804bp1805bp和1806bp處TC在秦川牛突變型個(gè)體中還在1700bp處C“T，而中國(guó)荷斯坦牛突變型個(gè)體在1638bp處插入了堿基Ga秦川牛和中國(guó)荷斯坦牛在1804bp.1805bp和1806飾處均為C堿基的研究結(jié)果目前尚無(wú)文獻(xiàn)報(bào)道，屬首次研究發(fā)現(xiàn)。2.FSHp基因第3外顯子及序列分析FSHp基因第3外顯子在秦

5、川牛上具有多態(tài)性，而在中國(guó)荷斯坦牛上未發(fā)現(xiàn)多態(tài)。在檢測(cè)的秦川牛群體中發(fā)現(xiàn)了兩種AA和BB兩種基因型。秦川牛在該位點(diǎn)群體多態(tài)信摘要3THERELATIONANALYSISOFTHETWINNINGPOLYMORPHISMOFFSHPGENEINQINCHUANCATTLEANDCHINAHOSTEINABSTRACTQinchuancattleandChinaHolsteinwereusedasexperimentalpopulation

6、.TheFSHpgenewerestudiedusingPCRSSCPandanalyzedthegeneticvariationTherelationbetweengeneticsmarkersandthetwinningwereanalyzed.Theaimistoexploreperfectmarkersintwinningwhichcanprovidetheorybaseonbreedingincatle.theresultsw

7、erefollows:1.Theanalysisonpolymorphismandsequenceonthe5flankofFSHpThepolymorphismwerefoundinPCRamplificationregionofthepremierFSH日P1FSHPP2andFSH3P3inQinchuancatlebutonlyonitsofthepremierFSH(3P2andFSHP43inChinaHolstein.(1

8、)ThreegenotypeAABBandABwerefoundinPCRamplificationregionofthepremierFSH(3PIinQinchuancatlebutonlyonegenotypeinChinaHolstein.Thepolymorphisminformationcontent(PIC)valuesweremoderatepolymorphism(0.5P025)whichmeanslargevari

9、ationintheloci.Sequencealignmentfortheregiondemonstratedthattherewerealotofmutation.Atposition65bpaddedGintheAAgenotypetherewerethreemutationwiththeposition73bp(addedT)281bp(1’一G)and284bp(GA).(2)Bothnormalandmutationalge

10、notypewerefoundinPCRamplificationregionofthepremierFSHPP2inQinchuancattleandChinaHolstein.Sequencefortheregiondemonstratedthattherewerealotofmutation.Therewereonemutationwith642bp(GA)innormalgenotypeandtwomutationwith642

11、bp(GA)and690bp(CG)inmutationalgenotypeinQinchuancatle.Therewereonemutationwith642(GA)innormalgenotypeandtwomutationwith642bp(GA)and642bp(CG)inmutationalgenotypeinQinchuancattle.(3)Bothnormalandmutationalgenotypewerefound

12、inPCRamplificationofthepremierFSHdemonstratedPP3inQinchuancattleandChinaHolsteinSequenceforregionthethattherewerealotofmutation.Therethreemutationlociregtonwith1804bp1805bpI806bpwhichthreeTchangedCinnormalgenotypeandmuta