Morozov G, Verlinsky O, Rechitsky S, Ivakhnenko V, Goltsman E, Gindilis V, Strom C, Kuliev A, Verlinsky Y
Reproductive Genetics Institute, Chicago, Illinois 60614, USA.
J Assist Reprod Genet. 1999 Apr;16(4):212-5. doi: 10.1023/a:1020368908134.
Because stage-specific genetic expression in human preimplantation development is not sufficiently studied, we have undertaken the construction of a subtraction complementary DNA (cDNA) library enriched for transcripts specific for human blastocysts.
For this purpose individual pools of cDNAs synthesized from four hatched blastocysts and three cleaving 8- to 10-cell embryos were exposed to suppression subtractive hybridization to minimize the presence of transcripts of housekeeping genes and other genes of maternal origin known to be expressed earlier in preimplantation development. Random clones of this library were sequenced and analyzed using the BLAST algorithm.
The resulting subtraction library had a complexity of 3 x 10(5) and an average size of inserts of about 0.8 kb. Sequencing of random library clones revealed the following human genes: CD9 antigen, fatty acid binding protein, ferritin heavy chain, amyloid precursor, MAP kinase messenger RNAs, DNA clone 127H14, messenger RNA for diacylglycerol kinase, a sequence homologous to C1 inhibitor, messenger RNA for the KIAA0145 gene, and others.
The presence of these genes in human preimplantation development suggests expression specific to the blastocyst stage.
由于人类植入前发育阶段特异性基因表达尚未得到充分研究，我们构建了一个消减互补DNA（cDNA）文库，该文库富含人类囊胚特异性转录本。
为此，将从四个孵化囊胚和三个处于8至10细胞期分裂胚胎中合成的单个cDNA池进行抑制消减杂交，以尽量减少管家基因转录本和已知在植入前发育早期表达的其他母源基因的存在。对该文库的随机克隆进行测序，并使用BLAST算法进行分析。
所得消减文库的复杂度为3×10⁵，插入片段平均大小约为0.8 kb。对文库随机克隆的测序揭示了以下人类基因：CD9抗原、脂肪酸结合蛋白、铁蛋白重链、淀粉样前体、MAP激酶信使RNA、DNA克隆127H14、二酰基甘油激酶信使RNA、与C1抑制剂同源的序列、KIAA0145基因信使RNA等。
这些基因在人类植入前发育中的存在表明它们在囊胚阶段特异性表达。
