Uses of MRPINK in restraining liveness of Macrobracium rosenbergii spermatozoon
A technology of Macrobrachium rosenbergii and sperm, applied in the application field of MRPINK in inhibiting the sperm activity of Macrobrachium rosenbergii, can solve the problems of zero understanding and operability
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Embodiment 1
[0017] Example 1: Obtaining of MRPINK gene and its encoded protein
[0018] For details, please refer to the literature: Cao JX, Dai JQ, Dai ZM, Yin GL and Yang WJ. Amale-reproduction-related kazal-type peptide inhibitor gene in the prawn, Macrobrachium rosenbergii: molecular characterization and expression patterns. Marine Biotechnology. 2007, 9: 45~55.
[0019] ① Take the male and female reproductive system tissues of Macrobrachium rosenbergii, and extract total RNA according to the instructions of the Trizol kit;
[0020] ②Establish the SSH cDNA library of the male reproductive system of Macrobrachium rosenbergii, and screen the differentially expressed gene fragments:
[0021] A. Synthesis of the first strand: Extract PolyA according to Oligotex mRNA Kits method + RNA to PolyA + RNA is used as a template for reverse transcription to synthesize single-stranded cDNA.
[0022] B. Synthesis of double-stranded (ds cDNA), digested with Rsa I enzyme to generate blunt ends.
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Embodiment 2
[0055] Example 2: Inhibitory effect of MRPINK on the sperm activity of Macrobrachium rosenbergii
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