本研究主要目的是針對與產蛋關係密切之卵巢進行抑制性雜合扣除法（SSH）分析卵巢中與產蛋性能相關之基因。經由雜交抑制扣除法所得高產品系扣除低產品系所得抑制性雜合扣除選殖株769個包含有59個已知基因、12個已發表之EST序列與16個未知基因；而低產品系扣除高產品系所得抑制性雜合扣除選殖株823個包含有26個已知基因、5個已發表之EST序列與7個未知基因。針對出現頻率較高的九個選殖基因，以半定量RT-PCR進一?確認其差異性表現，結果發現EF1A1、mitochondrion 5、mitochondrion 6、Hsp70、FHCP、Unk-9和Unk-10基因在高產蛋率品系表現量比低產蛋率品系表現量高，其中mitochondrion 5和Hsp70基因達顯著差異；GTPase TC10和GST基因在低產蛋率品系表現量比高產蛋率品系表現量高，其中GST基因達顯著差異。 針對選殖所得差異性基因土雞卵巢Unk-9基因進行功能性分析，結果發現所得之序列片段為similar to hypothetical protein FLJ22344 (LOC427112) 3’端序列，與雞隻基因體作比對，可得知Unk-9位於雞隻性染色體W上，此基因具有ASN glycosylation site、CK2 phosphorylation site及兩個PKC phosphorylation site，但未能分析到有任何的功能性domain，故對其作用仍一無所知。 利用西方點墨法針對土雞卵巢Hsp70蛋白質表現做探討，結果顯示Hsp70蛋白質在高產蛋土雞卵巢組織表現量顯著高於低產蛋土雞。然而，Hsp70蛋白質在高產蛋土雞及低產蛋土雞肝臟及肌肉組織表現則無差異，推論高產蛋土雞可能藉由較高的Hsp70蛋白質表現量影響土雞的生殖性能。 綜合以上結果顯示，利用抑制性扣除法可選殖出高產蛋品系及低產蛋品系卵巢差異性基因，而這些差異性基因在產蛋機制所扮演的角色仍有待進一?的探討。 The goal of this study was to identify the differential expression of genes in the ovaries of high and low egg production lines from Taiwan country chicken. By using suppression subtractive hybridization method (SSH), Fifty nine known genes, 12 EST clone sequences and 16 unknown genes were identified from 769 SSH clones of high egg production line subtracted low egg production line. In addition, twenty six known genes, 5 EST clone sequences and 7 unknown genes were identified from 823 SSH clones of low egg production line subtracted high egg production line. To investigate the involvement of these genes in egg production, nine genes with high expression level were selected to confirm the differential expression by RT-PCR. The results showed that elongation factor 1A1, mitochondrion 6, ferritin heavy chain protein, as well as Unk-9 and Unk-10 of the unknown genes expressed tended to be higher in high egg production line than those in low egg production line while mitochondrion 5 and Hsp70 gene expression in high egg production line were significantly higher than those in low egg production line. GTPase TC10 gene expression in low egg production line tended to be higher than that in high egg production line, whereas expression of GST gene in low egg production line was significantly higher than that in high egg production line. To further analyze the possible function of Unk-9 gene in ovaries, cDNA sequences were blasted in NCBI. The result showed that this sequence is homologous to the 3’ end of similar to hypothetical protein FLJ22344 (LOC427112). This gene located at sex chromosome W in chicken. It contains one ASN glycosylation site, one CK2 phosphorylation site and two protein kinase C phosphorylation sites. However, there is no functional domain has been found in this protein and the function of this protein remained unclear. The protein levels of Hsp70 in high and low egg production lines were analyzed by Western blot analysis. The result showed that Hsp70 protein level of high egg production line was significantly higher than that of low egg production line in ovaries. However, no significant difference was found in skeletal muscle and liver of high and low egg production lines. These results imply that Hsp70 may be involved in the egg production in ovary.
