[Cloning, expression, and antibody production of mouse ISP2].
A cDNA encoding mouse implantation serine proteinase 2   ISP2  was amplified from total cDNAs of mouse uterus implantation sites on D4.5 of pregnancy by PCR, and sequenced  GenBank accession No. A442918  . DNA sequencing indicated that the ISP2 cDNA had an unreported 204 bp sequence at 3' untranslated region besides the open reading frame encoding 279 amino acid residues, which was identical with literature. In order to obtain recombinant ISP2  rISP2     an expression plasmid pGEX-4T-2/ISP2 was constructed and transformed into E.coli BL21(DE3) strain. Expressed fusion protein GST-ISP2 was purified by SDS-PAGE and digested with thrombin, and the digestion mixture was subjected to SDS-PAGE again to recover rISP2. Rabbits were immunized using rISP2 as immunogen, and the polyclonal anti-ISP2 antisera were obtained. Immunohistochemical analysis using this antisera showed specific and high expression of ISP2 in mouse endometrial gland epithelium in early pregnancy.
Huang
Zhe-Ping
ZP
National Laboratory of Contraceptive and Devices Research, Shanghai Institute of Planned Parenthood Research, Shanghai 200032, China.
Wang
Jian
J
Yu
Hao
H
Shen
Wei-Xiong
WX
Huang
Ping
P
Tso
Jia-Ke
JK
Yang
Zeng-Ming
ZM
Shen
Qing-Xiang
QX
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D003001:Cloning, Molecular; D004926:Escherichia coli / Q000235:genetics; D005260:Female; D007150:Immunohistochemistry; D008297:Male; D051379:Mice; D008807:Mice, Inbred BALB C; D008969:Molecular Sequence Data; D010957:Plasmids; D011817:Rabbits; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis / Q000276:immunology; D012697:Serine Endopeptidases / Q000737:chemistry / Q000235:genetics* / Q000276:immunology
D011993:Recombinant Fusion Proteins; C436409:Prss29 protein, mouse; D012697:Serine Endopeptidases