; L 0254 H1023 , State Key Laboratory of Reproductive Biology, Institute of Zoology, The Chinese Academy of Sciences, Beijing , China

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1 50 (3) : , 2004 A cta Zoologica S inica G0101C0323 L 0254 H , ,, ( ESTs) G0101C0323 L 0254 H1023 EST, Northern :,, ; G0101C %, L 0254 H1023, ; G0101C0323, ; L 0254 H1023 [ 50 (3) : , 2004 ] G0101C0323 L 0254 H1023 Differential expression of G0101C0323 and L 0254 H1023 in mouse uterus during the es2 trous cycle 3 L I Fei2Xue, TAN Yin2Fei, WAN G Yan2Ling 33 State Key Laboratory of Reproductive Biology, Institute of Zoology, The Chinese Academy of Sciences, Beijing , China Abstract Temporal and spatial change of two differential express sequence tags ( EST), G0101C0323 and L 0254 H1023, in cycling mice uteri were studied by Northern blot analysis and in situ hybridization. It was demonstrated that the ex2 pression level of G0101C0323 and L 0254 H1023 was higher at the diestrous stage and much lower at the estrous stage. The expression of G0101C0323 at the estrous stage was 27 % of the expression at the diestrous stage. The expression of L 0254 H1023 at the estrous stage could not be detected. In situ hybridization further showed that the transcript of G0101C0323 was mainly localized in the luminal and glandular epithelium during the estrous cycle, whereas L 0254 H1023 in uterine stromal cells. The data indicated that the genes might participate in some cell events such as apoptosis in mice u2 teri under the regulation of ovarian steroid hormones [ Acta Zoologica Sinica 50 (3) : , 2004 ]. Key words Estrous cycle, Differential express gene, G0101C0323, L 0254 H1023,, ;, 4-5,,, ;,,, (Burroughs et al., 2000), ; , ( G ) ( KSCX222SW2201 KSCX32IOZ207) [ This research was funded by the grants from the State Key Basic Research Program (No. G ) and Chinese Academy of Sciences (No. KSCX2 22SW2201, KSCX32IOZ207) ] 33 (Corresponding author). ν 2004 Acta Zoologica Sinica E2mail :

2 ,, 111, CD21 (5-6, g), 25 ( Tan et, 12L 12D,, al., 2003), 62, ( Nelson et al., DNA 1982) 40, ( ESTs), EST - 80 cdna, 112 Northern EST, L 0254 H1023 (1), G0101C0323 L 0254 H1023 (815 ) (319 ) cdna, p GEM2T easy, G0101C0323 ( Promega), ( PPIase) 2 ( Ppil2) ; Prime2a2gene system ( Promega) PPIase cdna [ 2 32 P ] dctp (, ), ;, nick ( Pharmacia) Northern (Renoir et al., 1994 ; Liou et al., 2002) L 0254 H1023 I GF21 RNA, 30g RNA 6 EST ( 1 %),, 1 NCB I G0101C0323 RNA, RT2PCR, Trizol ( Gibco BRL) Hybond + ( Pharmacia), 8 s (012 mol p H 714, 011 G0101C0323 L 0254 H1023 mmol ED TA, 7 % [ w/ v ] SDS, 1 % [ w/ v ] BSA,, 15 % [ v/ v] ) 65 4 h, 32 P ;, X ( 1 PCR Table 1 PCR primers sequences of genes ( Gene) ( Primers sequence) ( Product size) (bp) Northern blot G0101C0323 F : 5 2 GTC CCA ACC TTT CAC ACA CG 824 R : 5 2 TCA CCA AGA ACA GCA GGA CC L 0254 H1023 F : 5 2TTC TCC AAA GAC TAT CAA GTG 432 R : 5 2CAA ACA GTA AAG GGG AAA GA PD H F : 5 2ACG ACC CCT TCA TTG ACC 700 R : 5 2TCA GAT GCC TGC TTC ACC ( In sit u hybridization) G0101C0323 F : 5 2 GAC TTC AGC TCC CAC CTC G R : 5 2 TCA CCA AGA ACA GCA GGA CC 339 L 0254 H1023 F : 5 2TTC TCC AAA GAC TAT CAA GTG 432 R : 5 2CAA ACA GTA AAG GGG AAA

3 3 : G0101 C0323 L 0254 H G0101C0323 A. G0101C0323 B : GAPDH G0101C0323 DiePro Est Met Fig11 Expressiom of G0101C0323 in cycling mice uteri A. Expression of G0101C0323 during the estrous cycle in the uterus. B. Comparative expression level of G0101C0323 during the es2 trous cycle in the uterus after normalized with that of GAPDH. Die/ 1, Pro/ 2, Est/ 3 and Met/ 4 means the stage of diestrous, proestrus, estrous and metestrus., ),,, Clearmount (MD, Amersham Biosciences, ), (Zymed) GA PD H 113 NCB I G0101C0323 L 0254 H1023 (1), 3,, RNA, RT2PCR cd2 3 ; Northern NA, cdna Q IAquick Spin ( Q IA2 GEN Inc. ), p GEM2T easy (Promega) S al Ncol ( Promega), Q I2 Aquick Spin ( Q IA GEN Inc. ), DIG2RNA ( ENZO DI2 A GNOSTICS. Inc) Northern G0101C0323, 011g/ l Qin et al. (2003),, 2 SSC %27 %38 %, K ( GIB ICO Inc. ), (1) t, P 4 % 015 % ; 01004, (50 %, 20 mmol Tris2HCl, 50 mmol ED TA, t RNA Coli 015 mg/ ml, D TT 100 mmol) 58 4 h, ; h SSC, Northern (2) 015 % (Bochringer Mannheim), (500 ) Northern L 0254 H1023, NB T BCIP (Bochringer Mannheim) 115 Northern GA PD H, S D, Studentπs t2test, P < G0101C kb,,, G0101C0323, 212 L 0254 H1023 mrna 715 kb ;

4 G0101C0323 DiePro Est Met Sense Fig12 Localization of G0101C0323 in cycling mice uteri Die, Pro, Est and Met mean the stage of diestrous, proestrus, estrous and metestrus. Sense means the negative control. 3 L 0254 H1023 A. L 0254 H1023 B. GAPDH L 0254 H1023 DiePro Est Met Fig13 Expressiom of L 0254 H1023 in cycling mice uteri A. Expression of L 0254 H1023 during the estrous cycle in the uterus. B. Comparative expression level of L 0254H1023 during the estrous cycle in the uterus after normalized with that of GAPDH. Die/ 1, Pro/ 2, Est/ 3 and Met/ 4 mean the stage of diestrous, proestrus, estrous and metestrus.

5 3 : G0101 C0323 L 0254 H L 0254 H1023 DiePro Est Met Sense Fig14 The localization of L 0254 H1023 in cycling mice uteri Die, Pro, Est and Met mean the stage of diestrous, proestrus, estrous and metestrus. Sense means the negative control., ;, 45 %66 % ( 3) t, P = 01002, L 0254 H1023,,,,,, (4), Northern ;, 3 L 0254 H1023 G0101C0323 EST,, (Jimmie et al., 2001), (V EGF) EST,, (Scholz et al., 1998 ; Mueller et

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