中國畜牧學會會誌33(增刊)  ,2004
J. Chin. Soc. Anim. Sci. 33(Suppl.)

119

豬粒線體DNA D-loop高變異序列定序分析

顏念慈(1) 簡嘉瑩(1) 林德育(1) 廖仁寶(1) 賴永裕(1) 張秀鑾(2) 吳明哲(1)

(1)行政院農業委員會畜產試驗所 (2)國立屏東科技大學

為探討豬隻粒線體DNA D-loop區域的多態性,本試驗設計了一組引子可以藍瑞斯、約克夏、杜洛克、盤克夏豬、挴山豬及桃園豬種豬之基因組DNA為模板,增殖出包含豬粒線體DNA D-loop區域的DNA片段,然後應用設計好之兩組適當引子對進行定序反應,以獲得DNA序列。將試驗所得D-loop區域DNA之序列與GenBank 編號AF034253序列比對,結果發現 (1)72頭種豬之D-loop區域位置1-710之序列共有17個位置發生變異,其中有16個變異為同類置換,初步可分成7種類型。(2)在序列位置711後有5’-CGTGCGTACA 10 bp的重複序列16-27個。(3)在位置711-1175之間與比對序列有4處位置與7處位置變異兩類型。綜合以上結果顯示DNA序列分析可產生較多之多態性。

關鍵語:豬粒線體DNA、D-loop區域、定序分析。

 

SEQUENCE ANALYSIS OF MITOCHONDRIAL DNA D-LOOP
HYPERVARIABLE REGION IN SWINE

N. T. Yen(1), C. Y. Chien(1), D. Y. Lin(1), R. B. Liaw(1), Y. Y. Lai(1), H. L. Chang(2), M. C. Wu(1)

(1) Livestock Research Institute, Council of Agriculture, Executive Yuan
(2) National Pingtung University of Science and Technology

Polymorphism of mitochondrial DNA (mtDNA) D-loop region in swine was studied, a pair of primers were designed to amplify the whole D-loop region of pig mtDNA from the genomic DNA of Landrace, Yorkshire, Duroc, Berkshire, Meishan and Taoyuan stock, then the PCR products were amplified by four suitable and designed primers. According to the reference sequence GenBank AF034253, there were 17 site substitutions in the sequence position 1 to 711 of D-loop region among 72 stock swine, sixteen nucleotide variations were transition. Seven type substitutions were distinguished. The number range of tandem repeat 5’-CGTGCGTACA after position 711 of D-loop region was 16 to 27. There were 4 site and 7 site substitutions in the sequence position 711 to 1175 of D-loop region. The results showed that DNA sequence analysis might distinguish more polymorphisms of swine.

Key Words: Swine mitochondrial DNA, D-loop region, Sequence analysis.