家族性高甘油三酯血症二例的临床与基因突变研究

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2 Silver Marten

3 . abcdp

4 abcdp

5 1. a 2 ch agouti Melanocytestimulating hormone, MSH MSH agouti

6 agouti Agouti agouti

7 agouti agouti agouti A non-agouti a

8 2. b 4 ch brown B b

9 2. b non-agouti C57BL/6

10 3. c 7 ch albino tyrosinase Albino C c albino

11 cc BALB/c AAbbcc

12 4. d 9 ch dilute D d

13 DBA/2J The Dilute Brown and non-agouti (DBA) strain aabbccdd

14 Dilute d, Nondilute D Brown b, Black B Nonagouti a, agouti A Albino c, nonalbino C Based on the earlier coding used by Little, Lane theorizes that the upper case "C" may have stood for the nonalbino allele.

15 7 ch pink-eyed dilution p Pink-eyed dilution non-agouti pinkeyed dilution

16 BALB/c AAbbcc C57BL/6J aabbcc A B C D A bbc D aab C D aabbc D ----cc --

17 Materials and Methods Mice BTLA-/- (10), PD-1-/- (17), and dual BTLA-/- and PD-1-/- mice were backcrossed for more than eight generations on a C57BL/6 background; HVEM-/- mice were generated by homologous recombination and backcrossed more than five generations on a B6 background (Q. Ye et al. manuscript in preparation). Wild-type C57BL/6 (H- 2b), BALB/c (H-2d), C57BL6/DBA F1 (H-2b/d), Bm12 (B6.C-H-2bm12/KhEg), and Bm1 (B6.C-H- 2bm1/ByJ) mice were purchased from The Jackson Laboratory, housed in specific pathogenfree conditions, and used for studies approved by the institutional animal care and use committee of Children s Hospital of Philadelphia.

18 . Species Stock Strain

19 1

23 AKR, C3H 7,8 97% ICR F1

24 2 20

25 Generation 0 Generation 1 G a / G a G b / G b G a / G b Generation 2 G a / G a G a / G b G b / G b Expected fractions ¼ ½ ¼ Incrosses of the types G a / G a G a / G a or G b / G b G b / G b types:crosses Other backcross intercross incross

26 N P 0 0 1/8 7 /32 53 / / / / /32768 =74.8% 98.4% N P 21st G 1.04% DBA,1909 C57BL,1921

27 . International Committee on Standardized Genetic Nomenclature for Mice Chairperson: Dr. Janan T. Eppig ( G.D. Snell is the first Chairman,1949 Chairperson: Dr. Goran Levan (

28 Dw, dwarf; Hbb, hemoglobin -chain Lpl Lipoprotein lipase Lpl, LPL

29 H-1, H-2 H-2 a, H-2 b

30 J or Jax Icr Rl Jr Mcw Kyo The Jackson Laboratory Institute for Cancer Research Drs. W.L. and L.B. Russell, Biology Division, Oak Ridge National Laboratory, Tennessee John Rapp Medical College of Wisconsin Kyoto University

31 1. A, NC, AKR, BALB A: Dr. A. Dux. Strong, 1921 Bagg NC: Cv: Dr. Verne M. Chapman

32 2. C57BL, C3H C3H/St: 1920, Strong, Bagg albino DBA C3H, C, CBA, CHI, C12I

33 F A(F87) DBA(?+F10)

34 strains CB CBA A A strains D1 DBA/1 strains AK AKR strains D2 DBA/2 strains B C57BL HR HRS/J B6 C57BL/6 strains L C57L/J B10 C57BL/10 strains R3 RIIIS/J BR C57BR/CD J SJL C BALB/c strains SW SWR C3 C3H strains

35 C: Cold Spring Harbor Laboratory 57: female No. 57 (male No.52) BL: Black 6: Substrain J: The Jackson Laboratory C57BR, C57BL C57BL/1, C57BL/6, C57BL/6J, C57BL/6N, C57BR/cdJ, C57L C57

36 Dilute d, Nondilute D Brown b, Black B Nonagouti a, agouti A Albino c, nonalbino C Based on the earlier coding used by Little, Lane theorizes that the upper case "C" may have stood for the nonalbino allele.

37 black black leaden (grey) C57 Mice from The Jackson Laboratory black C57BL/6 Is now probably the most widely used of all black inbred strains. brown

38 origin C non-albino (aabbcc) 57 female 57 BL black substrains C57BL/6J C57BL/10J C57BL/10SnJ C57BL/6NJ C57BL/6JEiJ C57BL/10SxJ C57BL/6By C57BL/6ByJ C57BL/10ScNJ C57BL/10ScSnJ C57BL/10WtRkJ C57BL/6HaJ

39 Substrains

40 A/He: Walter Heston A A / WyDiSn: Woolley A Dickie Snell

41 BALB/c :1913 H. Bagg ; 1927 MacDowell Snell; 1932, Snell /c DBA/1 DBA/2

42 F1 Hybrids F1 F1. D2B6F1 DBA/2 C57BL/6J. (DBA/2N C57BL/6J) F1. B6D2F1 (C57BL/6J DBA/2N)F1.

43 Recombinant Inbred Strains RI. X CXB, CXB2, CXB3 CB1BD22F1 CXB1 BXD22 ; (CXB1/ByJ BXD22/TyJ)F1.

44 Coisogenic Strains -, Lpl 447, db, ob : ( ) DBA/Ha - D C3H/N - +/W v

45 Segregating Inbred Strains - WB-w/+ BALB/c-nu/ +

46 nu/nu nu/+ nu/nu nu/+ nu/nu nu/+ dy

47 Congenic Strains ( ). B H-12 b C57BL/10Sn (B10), 129/J

48 The Journal of Immunology Table H-2 Mutants H-2 Haplotype designation Strain designation bm1 B6.C-H-2 bm1 bm2 B6.C-H-2 bm2 bm3 B6.C-H-2 bm3 bm12 B6.C-H-2 bm12

49 K I-A S D B10(C57BL/10) H-2 b b b b b C57BL/6 H-2 b b b b b DBA/2 H-2 d d d d d BALB/c H-2 d d d d d B10.D2 H-2 d d d d d C3H H-2 k k k k k CBA H-2 k k k k k B10.BR H-2 k k k k k

50 BTLA -/- (10), PD-1 -/- (17), and dual BTLA -/- and PD-1 -/- mice were backcrossed for more than eight generations on a C57BL/6 background; HVEM-/- mice were generated by homologous recombination and backcrossed more than five generations on a B6 background (Q. Ye et al. manuscript in preparation).

51 Wild-type C57BL/6 (H-2 b ), BALB/c (H-2 d ), C57BL6/DBA F1 (H-2 b/d ), Bm12 (B6.C-H-2 bm12 /KhEg), and Bm1 (B6.C- H2bm1 /ByJ) mice were purchased from The Jackson Laboratory, housed in specific pathogen-free conditions, and used for studies approved by the institutional animal care and use committee of Children s Hospital of Philadelphia.

52 Materials and Methods Mice BTLA -/- (10), PD-1 -/- (17), and dual BTLA -/- and PD-1 -/- mice were backcrossed for more than eight generations on a C57BL/6 background; HVEM/ mice were generated by homologous recombination and backcrossed more than five generations on a B6 background (Q. Ye et al. manuscript in preparation). Wild-type C57BL/6 (H-2b), BALB/c (H-2d), C57BL6/DBA F1 (H-2b/d), Bm12 (B6.C- H2bm12/KhEg), and Bm1 (B6.CH2bm1/ByJ) mice were purchased from The Jackson Laboratory, housed in specific pathogen-free conditions, and used for studies approved by the institutional animal care and use committee of Children s Hospital of Philadelphia. Reagents An Armenian hamster

53 . A. 1. Uniformity 2. Individuality B. 3. Homozygosity 4. Genetic stability 5. Isogenicity 6. Identifiability C. 7. International distribution 8. Backgroud data availability

54 . Residual heterozygosity.

55 Inbred mice inbreeding depression F1 hybrid vigor

56 1. 2.

57 3

58 . 1. (embro stem cell, ESC) ES 3~5 inner cell mass, ICM 2.

59 1. E14TG2a 129/Ola : Agouti : AA c : c ch c ch, p : pp C C C ch C e c C>C ch >C e >c (C), (C ch ), (C h ) (c)

60 129/Ola, AAc ch c ch pp,. C57BL/6J aabbcc Chimeras, ESC

61 Chimeras

62 129/Ola mice, AAc ch c ch pp, Creamy-white colored with pink eyes. C57BL/6J aabbcc A B C D, A bbc D, aab C D, aabbc D, cc --,

69 2. Targeting vector a. Homologous arms ( ) b. (hprt hypoxanthine phosphoribosyl-transferase gene ), neomycin resistance gene, neo r, G418,, (HSV-tk, herpes simplix virus thymidine kinase), (Ganc )

70 b.

71 ① 正向选择基因 - neor

72 Double positivenegative selection genes PNS G418 G418 Ganc G418

73 . apoe 1. apoe apoe cdna apoe- (Sac I/Bgl II )

74 2. pjpb63, 7.8-kb EcoRI neo 1.1-kb Sac I 1 3.

75 3. pnmc109,, 7.8-kb EcoRI neo Xho I/ BamHI 3 3. HSV-TK 3'

76 4. PCR Southern blotting (d) Southern blot analysis of genomic DNA from parental (P) and targeted (T) cell lines. Fragment sizes are in kb

78 , A w,, c ch p,

79 6. C57BL/6J DNA

80 C57BL/6J,, aabbcc ESC A w c ch and p,

81 7. apoe apoe, -/-, immunoprecipitate +/- +/+,

(Microsoft Word - 2011\262\243\276P\260\312\252\253\260\267\261d\260\335\303D Q\241\256A 1000221 _2_.doc)

$(Microsoft Word - 2011\262\243\276P\260\312\252\253\260\267\261d\260\335\303D Q\241\256A 1000221 _2_.doc)$ 動物出貨一般客訴問題 :(2011 2011/02 修 ) Q1: 同週齡小鼠體重體型大小差距較大 A: 本中心出貨小鼠, 皆以週齡來計算, 同週第一天與最後一天往往體重會有一些差距, 出貨時, 會盡可能的讓同一箱的出貨動物体型較為均勻, 以期達