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HEREDITAS (Beijing) 2009 1, 31(1): 63 68 ISSN 0253-9772 www.chinagene.cn 研究报告 DOI: 10.3724/SP.J.1005.2009.00063 猪 PRLR 和 RBP4 基因多态性与产仔性能的关系 孙延晓, 曾勇庆, 唐辉, 樊新忠, 陈其美, 李华, 钱源, 宋一萍, 271018 采用 PCR-RFLP 方法, 对莱芜黑猪 鲁莱黑猪 里岔黑猪 鲁烟白猪 新沂蒙黑猪 5 个山东地方 / 培育猪种和大约克夏 长白 杜洛克 3 个引进猪种共 8 个猪种 323 头繁殖母猪进行 PRLR 和 RBP4 基因的多态性检测, 并采用最小二乘法分析其对产仔数影响的遗传效应 结果表明 : 两个基因位点在 8 个猪种的测定群体中均存在多态性, 但山东地方 / 培育猪种与引进猪种间在基因型频率上存在较大差异 PRLR 和 RBP4 基因对产仔数性状有显著影响 (P<0.05), AA 均为优良基因型 对于 PRLR 基因, 山东地方 / 培育猪种内 AA 基因型母猪的总产仔数和活产仔数比 BB 基因型母猪平均多产 1.03 头和 0.89 头, 引进猪种中 AA 基因型母猪比 BB 基因型母猪平均多产分别为 1.26 头和 1.11 头 对于 RBP4 基因, 山东地方 / 培育猪种内 AA 基因型母猪的总产仔数和活产仔数比 BB 基因型母猪平均多产 0.59 头和 0.51 头, 引进猪种中 AA 基因型母猪比 BB 基因型母猪平均多产分别为 0.72 头和 0.64 头 猪 ; PRLR 基因 ; RBP4 基因 ; 多态性 ; 产仔数 Relationship of genetic polymorphism of PRLR and RBP4 genes with litter size traits in pig SUN Yan-Xiao, ZENG Yong-Qing, TANG Hui, FAN Xin-Zhong, CHEN Qi-Mei, LI Hua, QIAN Yuan, SONG Yi-Ping College of Animal Science and Veterinary Medicine, Shandong Agricultural University, Tai an 271018, China Abstract: The polymorphism of prolactin receptor (PRLR) and retinol-binding protein 4 (RBP4) genes was detected by PCR-RFLP method, and the effects of PRLR and RBP4 genes on litter size traits in pig were analyzed by the least square analysis. Data of total number born (TNB) and number born alive (NBA) from 323 sows, including five Shandong local pig breeds and three foreign pig breeds, were collected. The results showed that the polymorphic sites of both PRLR and RBP4 genes were found in all populations tested. The genotype distribution, however, revealed great differences between Shandong local pig breeds and foreign pig breeds. The effects of genotypes on TNB and NBA were significant (P<0.05). The homozygote AA was the most prolific genotype. For PRLR gene, AA genotypic sows of Shandong local pig breeds and foreign pig breeds produced 1.03 TNB, 0.89 NBA, and 1.26 TNB, 1.11 NBA more than BB genotypic sows, respectively. For RBP4 gene, AA genotypic sows of Shandong local pig breeds and foreign pig breeds produced 0.59 TNB, 0.51 NBA and 0.72 TNB, 0.64 NBA more than BB genotypic sows, respectively. Keywords: pig; PRLR gene; RBP4 gene; polymorphism; litter size 收稿日期 : 2008 04 02; 修回日期 : 2008 06 15 基金项目 : ( 2007249051), ( 2007LZ013) ( J07WF08) 作者简介 : (1981 ),,, E-mail: yxsun@sdau.edu.cn 通讯作者 : (1964 ),,,, Tel: 0538-8196883; E-mail: yqzeng@sdau.edu.cn

64 HEREDITAS (Beijing) 2009 31,, [1],,,, (Prolactin, PRL),,,,,, (Prolactin receptor, PRLR) [2, 3] 1998, Vincent [4], PRLR (Retinol-binding proteins, RBPs) A A, A [5], RBP4, RBP4,, [6, 7],,,, 5 3, PCR-RFLP PRLR RBP4,, 1 1.1 5 : 36 48, ; 30, ; 30, ; 54, 3 : 49 51 25, 8 323 ( 1 276, 2 7, ),,,,, 1.2 1.2.1 基因组 DNA 提取 - DNA, 0.8%, DNA, 25 ng/μl, 20 1.2.2 引物设计 [8, 9] PRLR : F: 5 -CGTGGCTCCGTTTGAAGAACC-3 R: 5 -C TGAAAGGAGTGCATAAAGCC-3 ; RBP4 F: 5 -GAGCAAGATGGAATGGGTT-3 R: 5 - CTCGGTGTCTGTAAAGGTG-3 1.2.3 PCR 扩增 (1)PRLR : 20 μl, : 10 Taq buffer 2.0 μl; 2.5 mmol/l dntp Mixture 1.6 μl; 20 μmol/l 0.5 μl; 2.5 U/μL Taq DNA 0.4 μl; DNA 1.0 μl; 14.0 μl PCR : 94 4 min; 94 30 s; 58 30 s; 72 30 s, 32, 72 7 min 1.5%, UVP (2)RBP4 : 25 μl, : 10 Taq buffer 2.5 μl; 2.5 mmol/l dntp Mixture 2.0 μl; 20 μmol/l 0.5 μl; 2.5 U/μL Taq DNA 0.4 μl; DNA 1.0 μl; 18.1 μl PCR : 95 5 min; 94 30 s; 56 45 s; 72 45 s, 35, 72 7 min 1.5%, UVP

1 : PRLR RBP4 65 1.2.4 RFLP 分析 (1)PRLR : 20 μl, 10 U/μL Alu 0.4 μl, 10 L buffer 2.0 μl, PCR 10 μl, 7.6 μl, 37 5 h, 3.0%, (2)RBP4 : 20 μl, 10 U/μL Msp 1.0 μl, 10 T buffer 2.0 μl, 0.1% BSA 2.0 μl, PCR 8.0 μl, 7.0 μl, 37 8 h, 1.5%, 1.2.5 统计分析 2, PopGen 32, SAS 8.1 GLM(General linear model),, : Y = / ; : Pi=[2(ii)+(ij 1 )+(ij 2 )+ +(ij n )]/2N : Pi i ; j 1, j 2,, j n i 1 n ; : n 2 ( Oi Ei 1/ 2) χ = Ei: E, Oi:, n: 2 2.1 PCR 2.1.1 PRLR 基因 PCR 扩增结果和 PCR-RFLP 分析 PCR 1.5%, 163 bp ( 1A) PCR-RFLP PCR Alu, 3 : 85/59/19 bp 104/85/59/19 bp 104/59 bp, AA AB BB ( 1B) 2.1.2 RBP4 基因 PCR 扩增结果和 PCR-RFLP 分析 PCR 1.5%, 480 bp ( 2A) PCR-RFLP PCR Msp, 3 : 190/154/136 bp 190/154/136/125/75 bp 154/136/125/75 bp, AA AB BB ( 2B) i i 2 图 1 PRLR 基因 PCR 扩增 (A) 及 PCR-RFLP 检测 (B) 结果 1 5 6 8 9: BB ; 2~4 7: AB ; 10: AA ; M: 20 bp DNA ladder 图 2 RBP4 基因 PCR 扩增 (A) 及 PCR-RFLP 检测 (B) 结果 1 2 5: AB ; 3 6: AA ; 4 7: BB ; M: Marker I

66 HEREDITAS (Beijing) 2009 31 2.2 PRLR RBP4 1, PRLR B( 0.5 ), AB>BB>AA; A, AA BB (P<0.05), A χ 2, - (P>0.05) 2, RBP4 3 A( 0.6 ), AA BB (P<0.05) /, AB>BB>AA, B, AA BB (P<0.05), A χ 2, - (P>0.05) 2.3 PRLR RBP4 / 3 PRLR (P<0.05); RBP4 / (P>0.05), / (P<0.05) /, AA, /, PRLR AA BB 1.03 0.89, RBP4 AA BB 0.59 0.51, PRLR AA BB 1.26 1.11, RBP4 AA BB 0.72 0.64 表 1 不同猪种 PRLR 基因型分布和等位基因频率 * AA AB BB A B Laiwu 36 0.0833(3) 0.5833(21) 0.3333(12) 0.3750 0.6250 2.6942 Licha 54 0.1296(7) 0.5000(27) 0.3704(20) 0.3796 0.6204 0.1474 Lulai 48 0.1042(5) 0.5208(25) 0.3750(18) 0.3646 0.6354 0.6216 Yimeng 30 0.4000(12) 0.3667(11) 0.2333(7) 0.5833 0.4167 2.0745 Luyan 30 0.3333(10) 0.4000(12) 0.2667(8) 0.5333 0.4667 1.3667 Landrace 51 0.1569(8) 0.4706(24) 0.3725(19) 0.3922 0.6078 0.0266 Yorkshire 49 0.1837(9) 0.5510(27) 0.2653(13) 0.4592 0.5048 0.4816 Duroc 25 0.6400(16) 0.3200(8) 0.0400(1) 0.8000 0.2000 0.0109 *: χ 2 0.05(1) 3.84, χ 2 0.01(1) 6.63 Laiwu: ; Licha: ; Lulai: ; Yimeng: ; Luyan: ; Landrace:, Yorkshire: ; Duroc: 表 2 不同猪种 RBP4 基因型分布和等位基因频率 AA AB BB A B Laiwu 36 0.1389(5) 0.6111(22) 0.2500(9) 0.4444 0.5556 1.7986 Licha 54 0.1458(7) 0.6042(29) 0.2500(12) 0.4479 0.5521 2.1395 Lulai 48 0.2037(11) 0.5185(28) 0.2778(15) 0.4630 0.5370 0.0603 Yimeng 30 0.3333(10) 0.5000(15) 0.1667(5) 0.5833 0.4167 0.0040 Luyan 30 0.3000(9) 0.5333(16) 0.1667(5) 0.5667 0.4333 0.1432 Landrace 51 0.4706(24) 0.4118(21) 0.1176(6) 0.6765 0.3235 0.2456 Yorkshire 49 0.3673(18) 0.4898(24) 0.1429(7) 0.6122 0.3878 0.0222 Duroc 25 0.7600(19) 0.2000(5) 0.0400(1) 0.8462 0.1538 0.4993

1 : PRLR RBP4 67 表 3 山东地方 / 培育猪种和引进猪种总产仔数和活产仔数的最小二乘分析 TNB NBA TNB NBA / PRLR AA 11.23±0.41 a 10.32±0.53 a 11.78±0.37 a 11.14±0.29 a AB 10.69±0.35 b 9.89±0.19 a 11.44±0.25 a 10.87±0.39 a BB 10.02±0.58 c 9.28±0.61 b 10.93±0.51 b 10.40±0.62 b RBP4 AA 11.13±0.31 a 10.31±0.41 a 11.67±0.41 11.05±0.27 AB 10.81±0.29 ab 10.02±0.42 ab 11.46±0.38 10.89±0.21 BB 10.38±0.21 b 9.64±0.45 b 11.24±0.31 10.70±0.45 PRLR AA 11.27±0.41 a 10.13±0.56 a 12.05±0.25 a 11.16±0.31 a AB 10.66±0.27 b 9.56±0.35 b 11.58±0.43 a 10.79±0.57 a BB 9.79±0.19 c 8.81±0.31 c 11.02±0.37 b 10.27±0.29 b RBP4 AA 11.18±0.21 a 10.24±0.39 a 11.72±0.32 a 10.85±0.27 a AB 10.79±0.43 ab 9.93±0.34 ab 11.51±0.21 ab 11.66±0.53 ab BB 10.31±0.18 b 9.45±0.42 b 11.15±0.41 b 10.37±0.52 b (P<0.05) TNB: ; NBA: 3 PRLR,,, [8, 10] [11] PRLR 10 Alu,, AA>AB>BB Rothschild [12] PRLR / / 6 PIC,, PRLR 6 4, AA BB 1 Terman [13] PRLR, PRLR (P<0.01),, AA AB BB, [14],, AA BB 2.41 2.77 (P<0.01), AA BB 3.33 3.30 (P<0.01), /, PRLR AA BB 1.03 0.89 (P<0.05), AA BB 1.26 1.11 (P<0.05), RBP4 [15, 16], RBP4,, [6, 7] Rothschild [17] 6 1 300, RBP4 AA BB 0.50 ( ) 0.26 ( ) Drogemuller [8] AA 1~10 BB 0.35, /, RBP4 AA BB 0.59 0.51, AA BB 0.72 0.64 (P<0.05), PRLR RBP4,, PRLR RBP4, PRLR RBP4 ;,

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