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Microsatellite Cloning and Screening of the AFLP Markers Linked to the Sex Locus of Acipenser Schrenckii

Author: LiuCui
Tutor: SunXiaoWen
School: Harbin Normal University
Course: Biochemistry and Molecular Biology
Keywords: AFLP Microsatellite Gender -specific markers Amur sturgeon
CLC: S917.4
Type: Master's thesis
Year: 2011
Downloads: 51
Quote: 0
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Abstract


Of Amur sturgeon (Acipenser schrenckii) is a cartilaginous fish (Chondrichthyes) Acipenseriformes (Acipenseriformes) Sturgeon Branch (Aeipenseridac) sturgeon is a fish (Acipenser Linnaeus), a unique sturgeon Heilongjiang River, with individual large, long life, the survival rate high, less disease, fast growth is the most economic value of rare Chinese sturgeon fish, farmed very broad prospects. In this paper, the AFLP technique belong to the Siberian sturgeon breeding populations of genetic structure is analyzed, with 7 pairs of primers 24 Siberian sturgeon carried the AFLP analysis, each pair of primer amplification bit points in 24 3 9 ' , the total amplified 234 loci polymorphic loci 116 polymorphism ratio was 48.83%, the average Nei's genetic diversity index H 0.4297 average Shannon's index I 0.1233 individuals The average genetic distance of 0.1872, indicating that the relatively low level of genetic diversity in the Siberian sturgeon farming groups. Magnetic beads enriched build Amur sturgeon partial genomic microsatellite library to radioisotope [γ 32 P] labeled probe (GA) 12 (CAT ) 8 , (AGAT) 6 hybrid screening of clone 3320 election 1200 sent sequencing, successful sequencing of 1136, 1048 (positive rate 92.25%) clones containing microsatellite sequences. Microsatellite loci 1529, the number of repeat microsatellite in more than 10 accounted for 65.24%. Perfect type of 1311 (85.74%), non-perfect 55 (3.60%), mixed in 163 (10.66%). Based on the sequence of both sides of the wing of the microsatellite repeat sequence, 3.0 of primers were designed using the primer design software Primer 299 pairs, Synthesis 200 pairs, Amur sturgeon DNA as a template for PCR amplification, screening results 100 pair of primers can be better amplified specific bands found including 16 pairs of primers can be amplified polymorphic product, can be used as genetic markers. Also screened two female specific molecular to mark HLJSX178 and HLJSX209 the research and development of microsatellite markers in the analysis of genetic variation, protect Biology and germplasm to a wide range of applications. High-salt and phenol chloroform isoamyl alcohol (PCI) method to extract DNA, using amplified fragment length polymorphism (amplified fragment length polymorphism, AFLP) molecular markers with 8 EcoR I-NNN and eight Mse I-NNN composition 64 pairs of primers, genomic DNA polymorphism detection Amur sturgeon male and female, screening Amur sturgeon gender-related molecular markers. The experiments were amplified 3264 with screening out the the candidate differences between DNA fragment 379. The present study is the first attempt to in Amur sturgeon genome find a gender-specific marker. Although not find such a marker, these data provide a further study of the relevant aspects of the useful information.

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CLC: > Agricultural Sciences > Aquaculture, fisheries > Aquatic basic science > Aquatic Biology > Aquatic Zoology
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