利用DNA池和测序技术快速筛查候选效应单倍型

选取产蛋性能具有明显差异的4个鸡品种（莱航鸡、阳山鸡、丝羽乌骨鸡和隐性白洛克鸡）构建品种DNA池，通过测序研究鸡催乳素基因5′-侧翼调控区部分序列（1028 bp）的单倍型，快速筛查到由4个多态位点（C-2402T、C-2161G、C-2062G和G-2040A）组成的2种最常见单倍型CCCG和TGGA，其中CCCG在莱航鸡品种中的频率接近1，可能利于产蛋。再利用RFLP和SSCP将这4个位点在品种及家系中进行基因分型，基因分型之后的统计结果与测序的结果基本相符。利用农大褐×泰和丝羽乌骨鸡家系检测CCCG单倍型的效应，发现含有此单倍型的单倍型组合具有更高的产蛋量。由6个位点组成的单倍型H3（CCTCTG，由CCCG再加上T-2101和T-2054组成）在莱航鸡的频率为1，在参考家系中检测H3的效应，发现含有H3的单倍型组合具有很高的产蛋量，并且与其它类型达到显著性差异。从本研究可见，利用这种方法能有效筛查到可能的效应单倍型。

Rapidly Screening Potential Causative Haplotype by DNA Pooling and Sequencing

Four breeds of chickens (White Leghorn, Yangshan, Taihe Silkies, White Recessive Rocks) with different egg production were applied to screen potential haplotype related to reproduction trait in part region of prolactin gene based on DNA pooling and sequencing. Two common haplotypes, CCCG and TGGA, of the most three ones CCCG, TGGA and TGCA which consisted with four SNPs sites (C-2402T, C-2161G, C-2062G and G-2040A) were found successfully in the fragment of 1028bp at the distal part of 5′flanking region. CCCG could be considered favourable for egg production since its frequency was estimated as 1 in White Leghorn which was verified by the results of PCR-RFLP and PCR-SSCP. Its effect on egg production was tested in the chicken reference of Nongdahe × Taihe Silkies, higher egg production was found in the haplotype combinations with CCCG, but no significant differences between them and the other haplotype combinations. The haplotype H3 (CCTCTG) which resulted from CCCG combining the other two sites (T-2101 and T-2054) was associated with egg production significantly in the chicken reference of Nongdahe × Taihe Silkies, and could be considered as a candidate marker for egg production. So rapidly screening potential causative haplotype could be succeeded applied DNA pooling and sequencing based on our researches.