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摘要：
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摘要：目的 建立介导胶原糖基化Glcα1、2Galβ1修饰基因（GLT25D1）敲低小鼠模型，为研究其在肝
脏疾病中的作用奠定基础。方法 基于Cre-loxp重组酶系统，采用胚胎干细胞线性打靶技术，获得同
源重组的干细胞，繁殖嵌合体，获得杂合子GLT25D1 +/- 小鼠。随后将GLT25D1 +/- 小鼠自交。通过PCR
凝胶电泳技术对子代小鼠的基因型进行鉴定。采用Western blot技术检测GLT25D1在WT型小鼠（wild
type，WT）和杂合子GLT25D1 +/- 小鼠各组织的表达。比较WT和GLT25D1 +/- 小鼠的生育情况、子代小
鼠基因型比例及出生20天的体重差异。结果 GLT25D1 +/- 小鼠配笼繁殖的264只子代小鼠中，杂合子
GLT25D1 +/- 小鼠152只，WT小鼠112只，无GLT25D1 -/- 小鼠。GLT25D1 +/- 小鼠数量和WT小鼠数量的比
值低于孟德尔遗传定律的2∶1（χ 2 = 9.818，P = 0.002）。Western blot示GLT25D1蛋白在WT小鼠的
心、肝、脾、肺、肾、脑和淋巴结等重要组织中均有表达，其中在肝、脾和肺中高表达，肾和淋巴
结中度表达，心和脑弱表达，GLT25D1 +/- 小鼠各组织表达水平均低于WT型小鼠。WT和GLT25D1 +/- 小
鼠在外观和行为上无显著差异，GLT25D1 +/- 小鼠出生后20天体重显著低于WT型小鼠（t = 2.520，P =
0.0177）。GLT25D1 +/- 小鼠交配后平均每窝出生的鼠仔数目为4只，低于WT小鼠平均鼠仔数7只（t =
-8.482，P ＜ 0.001）。结论 GLT25D1基因敲低后会导致GLT25D1 -/- 胚胎致死，且影响部分GLT25D1 +/-
小鼠正常出生，GLT25D1 +/- 小鼠出生后20天体重减轻，繁殖能力下降。
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Abstract: Objective To build the collagen glycosylated Glcα1 and 2Galβ1 modified gene (GLT25D1)
knockdown mice model to investigate its role in liver diseases. Methods Based on Cre-loxp recombinase system,
embryonic stem cell line targeting technology was implemented to obtain homologous recombination stem cells,
reproduce chimeras and obtain heterozygous GLT25D1 +/- mice. Then the heterozygote GLT25D1 +/- mice were
inbred and the genotype of offspring mouse was identified by PCR technique. The expression of GLT25D1
protein in WT mice (wild type, WT) and heterozygote GLT25D1 +/- mice in tissues were detected by Western
blot technique. The fertility of the WT and GLT25D1 +/- mice, the proportion of genotypes in the offspring mouse
and the weight at 20 days after birth were compared. Results Total of 264 mice were reproduced, including 152
GLT25D1 +/- , 112 WT and 0 GLT25D1 -/- . The ratio of the number of GLT25D1 +/- mice to the number of WT mice
was lower than that of Mendel’s genetic law 2∶1 (χ 2 = 9.818，P = 0.002). Western blot showed that GLT25D1
protein expressed in heart, liver, spleen, lung, kidney, brain and lymph nodes of WT mice, which were high
expressed in liver, spleen and lung, medium expressed in kidney and lymph nodes, and weak expressed in heart
and brain. The expression level of GLT25D1 in GLT25D1 -/- mice tissues was generally lower than that in WT
mice. There was no significant difference on the appearance and behavior of two genotype mouse. The weight
of GLT25D1 +/- mice at 20 days after birth was lower than that of the WT mice (t = 2.520, P = 0.0177). After
GLT25D1 +/- mating, the number of mice born per litter was 4, which was less than that of WT mice (t = -8.482，
P ＜ 0.001). Conclusions The GLT25D1 +/- mice showed a relatively low reproductive capability and abnormal
development. Compared with the WT mice, the weight of GLT25D1 +/- mice decreased.
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