跳到主要內容

臺灣博碩士論文加值系統

(44.200.77.92) 您好!臺灣時間:2024/02/25 03:00
字體大小: 字級放大   字級縮小   預設字形  
回查詢結果 :::

詳目顯示

我願授權國圖
: 
twitterline
研究生:曾文芳
研究生(外文):Tseng, Wen-fang
論文名稱:激濾泡素接受體基因於各豬種間之多態性
論文名稱(外文):The polymorphisms of follicular stimulating hormone receptor gene among defferent breeds of pigs
指導教授:鄭登貴鄭登貴引用關係吳和光---
指導教授(外文):Cheng Winston Teng-kueiWu Her-kuang
學位類別:碩士
校院名稱:國立臺灣大學
系所名稱:畜產學系研究所
學門:農業科學學門
學類:畜牧學類
論文種類:學術論文
論文出版年:1998
畢業學年度:86
語文別:中文
論文頁數:85
中文關鍵詞:激濾泡素接受體窩仔數梅山豬多產性限制脢截切多態性轉錄因子
外文關鍵詞:follicular stimulating hormone receptorlitter sizeMeishan pigpolificacyrestriction fragment length polymorphismGHF-1
相關次數:
  • 被引用被引用:0
  • 點閱點閱:136
  • 評分評分:
  • 下載下載:0
  • 收藏至我的研究室書目清單書目收藏:0
本研究首在針對包括梅山豬、藍瑞斯、約克夏、杜洛克、小耳豬、及桃園豬等,不同豬種
之激濾泡素接受體基因(follicular stimulating hormone receptor gene; FSHR gene)
,進行分析其核酸限制截切片段長度之多態性,並探討此等多態性與母豬多產性之關係
性;此外,試驗並亦針對前述各不同豬種,分別將其FSHR基因之上游序列部分進行選殖、
定序及比對,冀能從而瞭解該基因表現之調控序列,在各豬種間之異同,俾可提供未來作
為改進商業豬種生殖性能之依據。供本試驗用之基因組DNA,分別係取自福昌種豬場之藍
瑞斯、約克夏、杜洛克、與藍瑞斯及約克夏之雜交豬、購自商用肉豬場所飼養之LYD雜交
豬及桃園豬、台大農業試驗場所飼養之蘭嶼小耳豬、與自臺灣省畜產試驗所引進之梅山豬
等,經採取其外耳組織樣品萃取而獲得。供進行南方氏分析所需之探針有二:一為根據豬
激濾泡素接受體cDNA之已知序列,在其第10表現子(exon10)序列中設計適當引子,並以LY
D 雜交豬之基因組DNA 作為模板(template),經應用聚合聯鎖反應 (PCR),擴增一段長
度為876 bp之DNA片段,並命名為probe 2;另一為根據小鼠、鼠及人等之激濾泡素接受體
基因上游序列,擇取具有高同質性(homalogy)部分做為參考,設計上游引子,再配合前述
豬激濾泡素接受體cDNA 之exon 1序列,設計下游引子,並以梅山豬基因組DAN作為模板,
經PCR擴增一段長度為1112 bp 之DNA片段,並命名為probe1。激濾泡素接體基因之核酸限
制截切片段長度多態性分析,係將前述萃取自各不同豬種之基因組DNA,分別先應用10
種不同核酸限制,予以進行單一酵素截切後,再使用probe1 或probe2進行南方氏雜合
分析。試驗結果顯示,基因組DNA之經Bam HI、EcoRV、NdeI、KpnI、PvuII截切,並以pro
be1進行雜合反應者,與經HpaI、StuI、KpnI、EcoRI、PstI、DraI截切並以probe2進行雜
合反應者,在各豬種間並無多態性可言;惟桃園豬及杜洛克豬之基因組DNA,經EcoRI截切
並以probe1作為探針進行雜合反應者,與LYD 基因組DNA經KpnI片段並以probe2進行雜合
反應者,分別均顯示其激濾泡素接受體的基因截切片段電泳模式及長度,顯著異於其他受
測試之豬種者,顯示該基因在不同豬種間確有多態性存在之事實。此外,根據probe 1經
定序所得獲之序列,設計上游引子,並以FSHR基因之exon 1 部分序列做為下游引子,分
別針對藍瑞斯、約克夏、杜洛克、桃園豬及小耳豬等五種豬之FSHR基因之上游序進行擴增
,所獲得之片段及probe1(來自梅山豬)分別進行選殖及定序、比對,結果証明之激濾泡素
基因之上游序列,在六個不同豬種間雖具有高達95.5%之同質性,但在梅山豬者於 -697核
酸序列處有一段長5 bp之缺失現象。鑑於此一缺失序列適發生於類似GHF-1感應子(respon
se element)之位置,此係梅山豬所僅見之現象,是否此一序列缺失之突變足以影響梅山
豬激濾泡素接受體基因之表現,從而導致梅山豬有較佳之排卵數,並有利於梅山豬胚之存
活率,其間作用之詳細機制為誠值吾人進一步詳加探討。

Purposes of the present studies were to investigate the differences among Me
ishan, Landrace, Yorkshire, Duroc, Taoyuan, and Small Ear pigs in their polymo
rphism of genomic DNA coded for the follicular stimulating hormone receptor (F
SHR), and the connection between the above polymorphism and polificacy. Also,
the 5' flanking region fragments of FSHR of the six breeds were cloned, sequen
ced, and then compare the sequences which may influence the regulation of FSHR
gene expression. These results may be used foimprovement of commercial pig fe
rtility. Genomic DNA used in these experiments were extracted from the tissu
e explants of Landrace, Yorkshire, Duroc, and Landrace Yorkshire hybrid kept i
n Formosa Fortune Farm, Small Ear pig kept in the experiment farm of National
Taiwan University, LYD hybrid bought frome commercial farm and Meishan pig imp
orted from Taiwain Livestock Research Institute. Two DNA probes used in Southe
rn blot analysis of FSHR were amplified by polymerase chain rection (PCR) tech
nique. The 876 bp one was amplified from LYD FSHR exon 10 and named as probe 2
, primer pair used for amplification was derived from published pig FSHR cDNA
sequence. The 1112 bp one was amplified from Meishan pig FSHR 5' flanking regi
on and part of exon 1 and named as probe 1, the up stream primer used for ampl
ification was derived from the consensus sequence of FSHR 5' flanking region o
f mice, rat, and human, and the down stream primer was derived from pig FSHR e
xon1 sequence. Experimental results appeared that among the 10 restriction enz
ymes tested, there was no restriction fragment length polymorphism (RFLP) foun
d in genomic FSHR gene among the six breeds of pigs after they had been respec
tively restricted with Bam HI, EcoRV, NdeI, KpnI, and PvuII, then hybridized w
ith probe 1, and respectively restricted with HpaI, StuI, KpnI, EcoRI, PstI, a
nd DraI, then hybridized with probe 2. Whereas, FSHR genomic DNA of Taoyuan pi
g and Duroc pig were found to show different patterns form other breeds of pig
after digested by EcoRI and hybridized with probe 1. And FSHR genomic DNA of
LYD pig were found to show a different length of fragments after digested by K
pnI and hybridized with probe 2. These results indicated that there are differ
ence exist in genomic FSHR genes of different breeds of pigs. In the further e
xperiment, the 5' flanking region fragments of FSHR of Landrace, Yorkshire, Du
roc, Taoyuan, and Small Ear pigs were amplified by PCR technique with primer p
air derived from probe 2 sequence. And these five fragments and probe 2 were c
loned, sequenced, and analyzed. Although the homology of FSHR 5' region sequen
ce in the six breeds of pig is 99.5%, the one of Meishan pig has a five bp del
etion at site -697 when compared with other breeds of pigs. The site where the
deletion took place is a putative GHF-1 response element. Whether the deletio
n change the regulation of FSHR gene expression, and improve the ovulation rat
e and early embryo survival rate of Meishan pig still need further investigati
on.

QRCODE
 
 
 
 
 
                                                                                                                                                                                                                                                                                                                                                                                                               
第一頁 上一頁 下一頁 最後一頁 top