Sequence analysis of libraries from individual human blastocysts.
OBJECTIVE
It has recently become possible to construct cDNA libraries from individual human blastocysts to investigate the expression of embryonic genes in human preimplantation development. We have previously reported the expression of beta-actin, CD-59, and homeobox OCT-3 and identified almost-complete homology of sequences to human histone 3.1 and human ribosome protein S25. In the present paper, our further sequencing analysis of cDNA libraries from single human blastocysts is described.


METHODS
cDNA libraries were constructed from 13 blastocysts. Sequence analysis was performed in 120 clones from one of these cDNA libraries with fragments of 50 to 1000 bp. Their sequence identity was analyzed using the expressed sequence tag (EST) database.


RESULTS
The presence of two housekeeping genes, hexokinase I and serine/threonine phosphorylase, and four other ESTs was demonstrated, the identity of which, with particular gene expression in preimplantation development, has not yet been established.


CONCLUSIONS
The data demonstrate the usefulness of constructing cDNA libraries from individual human blastocysts and their values in the analysis of genetic expression in human preimplantation development.
Morozov
G
G
Reproductive Genetics Institute, Chicago, Illinois 60657, USA.
Verlinsky
O
O
Rechitsky
S
S
Kukharenko
V
V
Goltsman
E
E
Ivakhnenko
V
V
Gindilis
V
V
Strom
C
C
Kuliev
A
A
Verlinsky
Y
Y
D001755:Blastocyst / Q000737:chemistry*; D001923:Brain Chemistry / Q000235:genetics; D018076:DNA, Complementary / Q000032:analysis; D005333:Fetus; D015870:Gene Expression; D015723:Gene Library; D005819:Genetic Markers; D006593:Hexokinase / Q000235:genetics; D006801:Humans; D010190:Pancreatic Neoplasms / Q000235:genetics; D010749:Phosphoprotein Phosphatases / Q000235:genetics; D016133:Polymerase Chain Reaction; D017422:Sequence Analysis, DNA / Q000379:methods*
D018076:DNA, Complementary; D005819:Genetic Markers; D006593:Hexokinase; D010749:Phosphoprotein Phosphatases