利用MS-PCR及DNA定序檢驗盤克夏豬種的緊迫基因

廖仁寶(1) 呂宜瑾(2) 賴永裕(1) 劉錦條(1) 劉英明(2) 吳明哲(1) 張秀鑾(1)

(1)台灣省畜產試驗所 (2)國立中興大學動物系

盤克夏豬種於1996年自美國引進,飼養於台灣省畜產試驗所。計有243頭後裔被用來檢驗其緊迫基因,使用MS-PCR(突變點拆離式聚合玟s鎖反應)中的三個引子直接檢測Hal-1843突變點,該突變是由C突變為T,因此Hal-1843基因型為CC者是正常豬,TT者是突變豬,而CT者是攜帶突變基因的豬。直接電泳MS-PCR產物時,CC者為114bp片段,TT者為134bp片段,而CT者具有114和134bp兩個片段。具有CC、CT和TT的豬分別有71.2、27.6、和1.2%。豬心臟樣材取自11頭豬,並自心臟萃取DNA後,再以MS-PCR檢驗Hal-1843基因型,結果與血樣DNA的檢驗相吻合。因此把MS-PCR產物經純化、增幅和在自動定序儀予以定序。盤克夏豬含有Hal-1843的DNA片段經定序後,序列跟Fujii et al.(1991, Science 253: 448-451)之報告一樣。綜而言之,畜產試驗所的盤克夏豬群之緊迫基因頻率為0.15。

關鍵語:盤克夏、緊迫基因、定序。

 

PORCINE STRESS GENE OF BERKSHIRE BREED DETECTED BY
MS-PCR AND DNA SEQUENCING

R. B. Liaw(1), I. G. Li(2), Y. Y. Lai(1), C. T. Liu(1), Y. M. Liou(2), M. C. Wu(1) and H. L. Chang(1)

(1)Taiwan Livestock Research Institute
(2)Department of Zoology, National Chung Hsing University

Berkshire breed was imported from USA in 1996 and raised at Taiwan Livestock Research Institute. Two hundred and forty-three progenies were tested with three primers in MS-PCR (Mutagenically separated polymerase chain reaction) for point mutation from C to T at sequence position 1843 of porcine stress gene. Genotypes of Hal-1843 were designated as CC for the normal, CT for the carrier and TT for the mutant. Electrophoresis after MS-PCR, CC pigs had 114 bp band, TT pigs had 134 bp band and CT pigs had both 114 and 134 bp bands. Pigs with CC, CT and TT genotype were 71.2, 27.6 and 1.2%, respectively. Eleven pigs were slaughtered for collection of hearts and DNA was extracted from heart tissue individually. Samples of purified DNA were tested with MS-PCR and three genotypes of Hal-1843 were confirmed with blood sample. Then the product of MS-PCR was purified, amplified and used to analyze the nuclotide sequence in an automated DNA sequencer. The sequence of the DNA fragment including the Hal-1843 point mutation of the Berkshire pig was the same as described in the study of Fujii et al. (1991, Science 253: 448-451). In conclusion, the frequency of stress gene of Berkshire pig at the Institute was 0.15.

Key Words: Berkshire, Stress gene, Sequencing.