臺灣博碩士論文加值系統

本試驗以迷你衛星、RAPD與RAMPO DNA指紋技術比較鴕鳥、雞、小鼠、
山羊與豬之DNA指紋以明瞭種內或種間之差異。 試驗結果顯示：同樣使
用重複序列(TG)6探針下，以HaeIII限制切割鴕鳥與雞DNA所得之迷你衛
星DNA指紋態樣較之於以HinfI限制切割者可得較清晰之指紋，但小鼠與
山羊者則反之。豬DNA無論是以aeIII或限制HinfI切割所得之迷你衛星
DNA指紋態樣皆較不佳。本試驗選自OPAA、OPAO、OPAV、OPC及OPE等系列
共54種逢機引子，其中有51種(94％)引子可產生具多態性之鴕鳥、雞、小
鼠、山羊與豬之RAPD指紋；而鴕鳥、雞、小鼠、山羊與豬DNA能以重複序
列(TG)6探針建立具多態性之RAMPO指紋之引子數目各為26(48.1％)
、11(23.7％)、46(85.2％)、38(70.4％)及43(79.6％)。不同引子增殖同
種動物DNA所得之RAPD指紋皆不相同，且其產物經重複序列(TG)6探針雜交
後所得RAMPO指紋之差異甚大。在RAPD指紋圖上相同位置之環帶，其核
酸序列未必相同，且並非所有的RAMPO片段皆可在RAPD指紋圖上相同位置
處觀察到。 在種間比較方面，以HaeIII與HinfI限制切割鴕鳥、雞、
小鼠、山羊與豬DNA後與重複序列(TG)6探針雜交均能產生種間差異明顯的
迷你衛星DNA指紋。以51種逢機引子形成之RAPD與RAMPO指紋比較此此五種
動物，結果亦可顯示相同引子增殖各種動物所得之RAPD指紋其環帶之數目
、位置與長度分佈之範圍皆不相同，亦即種間之指紋態樣差異甚大，且隨
著引子之不同所得之指紋態樣亦不同；鴕鳥與雞DNA使用不同引子以RAPD-
PCR增殖，所得之產物大多無法與重複序列(TG)6探針雜合上。比較迷你衛
星、RAPD與RAMPO此三種指紋分析技術，迷你衛星DNA指紋技術之重複性較
高，但較之於RAPD與RAMPO指紋技術，卻較為費時、費力、費財且耗費DNA
量大，而RAPD與RAMPO指紋技術所需DNA量微又能產生種間差異明顯之DNA
指紋 ，因此單就比較種間之差異性，RAPD指紋足以勝任，而與RAMPO指紋
配合則可用以防止環帶之誤判。

Minisatellite, RAPD and RAMPO techniques were employed for
studying of fingerprints in ostriches, chickens, mice, goats and
pigs. They were then used for investigation the difference of
various species and inividual variations. The minisatellite
fingerprints of ostrich and chicken DNA digested with HaeIII and
hybridized with probe (TG)6 showed a better resolution as
compared to that by HinfI digestion. But it did not have a
good resolution in mouse and goat. The band pattern of
minisatellite fingerprints of pig DNA digested with either
HaeIII or HinfI restriction enzyme was shapeless. The
94％(51/54) of random primers in OPAA, OPAO, OPAV, OPC and OPE
serials can generated polymorphic RAPD fingerprints in ostrich,
chicken, mouse, goat and pig. Numbers(％) of primers which can
generated polymorphic RAMPO DNA fingerprints in oftrich,
chicken, mouse, goat and pig was 26 (40.1％), 11(23.7％),
46(85.2％), 38(70.4％) and 43(79.6％), respectively. The RAPD
fingerprints amplified with different primers had distinct
results, and the RAMPO fingerprint after hybridization from
RAPD products by probe (TG)6 exhibited differences among
individuals in the same species. The nucleotide sequence of the
bands at same position in the RAPD fingerprint coule be
different and the visual bands on the RAMPO fingerprint did not
observed in the RAPD fingerprint. Among species, the
minisatellite DNA fingerprints of ostrich, chicken, mouse, goat
and pig with HaeIII and HinfI restriction digestion and probe
(TG)6 hybridization were different extinctly. Numbers or lengths
of bands in RAPD and RAMPO fingerprints of various species were
different distinguishably when fingerprints were revealed with
the 51 primers. Most of DNA products of ostrich and chicken
amplified with different primers by RAPD-PCR did not hybridize
with probe (TG)6 . Comparison on fingerprints with methods of
minisatellite, RAPD and RAMPO, the minisatellite methods was
repeatable but more time-, labor-, cost- and DNA-consuming.
Based upon the fact that a less requirement os DNA amounts in
RAPD and RAMPO fingerprinting and generation of extinct banding
among species, the RAPD fingerprint was applicable for the
difference of various species. When both RAPD fingerprint and
RAMPO fingerprint methods were used conjunctly, it could avoid
misjudging of specified bands.