Abstract: Objective The expression level of peripheral hemoglobin aGal in the offspring of the Wuzhishan inbred min-Pig (WZSP) and its aGal knock gene pig offspring was carried out. Method The expression levels of peripheral erythrocyte aGal in the offspring of different generations of WZSP inbred lines and their GGTA1 / b4GalNT2 knockout pigs were measured by using the separation technology of fluorescein isothiocyanate ( FITC) . Result The average percentage of immunofluorescence response of aGal on the 26 heads red blood cells in the inbred line F_{27- 28} was 56. 17% ( 16. 2% - 86. 2%) . Among them, the average value of F₂₄ 8 heads red blood cells of aGal cells was 65. 39% ( 35. 5% -99. 2%) ,the average value of F_{1 9 - 21} 21 heads is 69. 67%(38. 8% -89. 4%) . The average values of the group group 1 and group 3 were significantly different ( P< 0. 05) . The further analysis showed that the percentage of immunofluorescence response of inbred red blood cells of aGal cells in inbred F_{27- 28} and F₂₄, F₂₄ and F₂₀. individual red blood cells of aGal cells decreased by 3. 07%, 4. 19% per generation; with the advancement of inbreeding lines, the immunofluorescence response of red blood cells of aGal cells decreased significantly. The immunofluorescence response of 8 offspring red blood cells of the knock gene modified pigs was 0. 1% which was consistent with the control group and could be regarded as homozygotes. It was with extremely low aGal expression was found, indicating that its immune rejection ability for xenotransplantation was low also. The study also found that the aGal expression of two siblings at F₂₇ WZSP was lower ( 16. 2% and 19. 5%) than that ( 31. 03%) of knock gene heterozygous pigs. Conclusion With the advancement of inbred lines, the expression of aGal in individual red blood cells was decreased gradually. Eight GGTA1 / b4GalNT2 knockout pigs were homozygous. The inbred line F₂₇ was an individual with low expression of aGal.

Key words: inbred pig, generation, gene, red cell, α-galactosyltransferase

摘要： 目的 探讨五指山小型猪近交系不同世代及其 GGTA1 / b4GalNT2 基因敲除猪繁育后个体外周血红细胞 aGal表达水平,为开发应用提供科学依据。 方法 利用异硫氰酸荧光素( FITC) 分离技术,对五指山小型猪近交系不同世代及其 GGTA1 / b4GalNT2 基因敲除猪繁育后代外周红细胞 aGal 表达水平进行了测定研究。 结果 组 1 近交系F_{27- 28}26 头小型猪红细胞 aGal 免疫荧光反应百分数平均值为 56. 17%(16. 2% ~ 86. 2%) ;组 2 F₂₄8 头小型猪平均值为 65. 39%(35. 5% ~ 99. 2%) ;组 3 F_{19- 21}21 头小型猪平均值为 82. 15%(38. 8% ~ 89. 4%) 。 组 1 与组 3 之间平均值差异有统计学意义(P<0. 05) ;进一步分析发现组 1 与组 2、组 2 与组 3 相比,个体红细胞 aGal 免疫荧光反应百分数代间分别下降 3. 07% 和 4. 19%。 组 4 中有 8 头 GGTA1 / b4GalNT2 基因敲除猪繁殖后代和 1 头已知为 GGTA1 / b4GalNT2+hCD46+hCD55+hTBM 五基因修饰纯合子猪的红细胞 aGal 免疫荧光反应测定值均为 0. 1%,与对照组结果一致。 参与测定的 8 头 GGTA1 / b4GalNT2 基因敲除猪繁育后代为纯合子个体。 同胞兄妹 F₂₇两头 aGal 低表达量为 16. 2%和 19. 5%,均低于 GGTA1 / b4GalNT2 基因敲除杂合子后代猪平均值 31. 03%。 结论 随近交系推进其个体红细胞 aGal 表达量逐步降低;8 头 GGTA1 / b4GalNT2 基因敲除猪繁育后代为纯合子个体;近交系 F₂₇两头为 aGal低表达个体。

关键词: 近交系猪, 世代, 基因, 红细胞, α 半乳糖转移酶